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最終更新日 : 2021年11月13日

iGEM2021全チームプロジェクト概要 ~第二弾~

本記事は、iGEM 2021年に参加した全チームのプロジェクトについてまとめた記事の第二弾になります。(全四回の第二回)。
iGEM 2021では、どのようなテーマがあったのか網羅的にわかるようになっておりますので、ざっと眺めて見てはいかがでしょうか。


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※本まとめは、全てのデータをiGEM2021のオープンなデータから取得しております。

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このような構成で、全チームをまとめていきます。

UTokyo
Title
YEAST-AID

Abstract
Incoming aging society asks us: "How can we extend the healthy life expectancy?". There are many obstacles to answering the question, one of which is bedsore. A bedsore is a severe wound that often occurs in people who spend a long time in bed. It requires continuous care by medical professionals and keeps patients away from home even after recovering from the initial symptoms. Our team suggests a solid solution to this problem by creating YEAST-AID for bedsore. This project aimed to help everyone takes care of bedsore correctly and efficiently, discharging millions of people from extended hospitalization. Our solution, YEAST-AID is a multifunctional wound dressing using synthetic yeast. The yeast has three functions supporting wound care: wound monitoring by oxygen concentration, infection prevention by antimicrobial peptides, and detection of P. aeruginosa infection. Furthermore, the yeast is appropriately integrated into the fiber-like hardware to function safely on the wound.
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BNU-China
Title
Cheat Me If You Can

Abstract
In industrial production, inevitable mutations and phenotypic variations will lead to the appearance of cheater cells which avoid the burden of synthesizing exogenous products. Given the liberation from synthesis, cheaters may gain remarkable survival advantages over common workers. Such strain degeneration will therefore lead to a great loss in both quality and quantity of the products. To solve this problem, BNU-China seeks inspiration from a natural colonial model, in which guard bacterial cells can specifically eliminate cheater cells to maintain the balance of the population. We decide to develop a universal cheater-proof platform for industrial production by introducing carefully engineered guards and workers into the system.
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TUDelft
Title
AptaVita: A novel and modular aptamer-based rapid diagnostic test for vitamin deficiencies

Abstract
Hidden hunger, synonymous with micronutrient deficiencies, is estimated to impact the health and life-quality of one in three people, especially in sub-Saharan Africa and South Asia. Current monitoring policies are insufficient, as more data on hidden hunger is required to implement solutions effectively. However, existing detection techniques remain inaccessible for most people. Here, we present AptaVita: a modular, quantitative, and accessible paper-based rapid diagnostic test for vitamin deficiencies available at the point-of-care. Aptazymes (ligand-regulated self-cleaving ribozymes) engineered through in vitro evolution for binding the vitamins of interest are fused with the lacZ reporter gene. Vitamin-responsive translation activation is enabled in the PURE cell-free expression system on paper support. The colorimetric output is quantified by a portable, 3D-printed, dedicated hardware, allowing us to assess the level of targeted vitamins. We envision AptaVita making hidden hunger visible and addressable, thereby contributing to monitoring and tackling vitamin deficiencies together with those involved.
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KU Leuven
Title
Development of a continuous directed evolution platform for in vitro-cultured plant cells

Abstract
Targeted continuous evolution, using a platform based on CRSIPR-Cas-based nickase and error-prone DNA polymerase (a.k.a. EvolvR), is a powerful approach to rapidly evolve specific phenotypes with minimal user intervention. By establishing this method in planta, we seek to accelerate the evolution of needed plant phenotypes such as (a)biotic stress tolerance. EvolvR, targeted to a gene of interest by a specific guide RNA, continuously diversify the underlying DNA, with cells constantly under selection for improved variants. We have confirmed the efficiency of the approach in E. coli, validated it in a plant-based cell-free system and confirmed its functionality in Tobacco BY-2 cell culture. We have also established mathematical approaches to model and optimize the selection regimens to maximize the efficiency of the pipeline. Our in silico, in vitro and in vivo results further demonstrate the potential of EvolvR to plant biotechnology improving crops and developing new plant-based applications.
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UCSC
Title
Progenie: A prokaryotic sequence-specific gene elimination system

Abstract
Shiga toxin-producing Escherichia coli (STEC) outbreaks are one of the leading causes of food recalls in the United States; infections can cause diarrhea, vomiting, fever, and even kidney failure and death. We developed a programmable in-situ gene-elimination system, Progenie, to selectively knock-out the Shiga-toxin producing gene stx2 from an environmental microbial community—reducing STEC outbreaks without harming commensal bacteria. Progenie consists of an initial phage delivery of a conjugatable plasmid encoding an RNA-guided transposon system that self-integrates into a targeted locus and spreads to neighboring microbes. Using E. coli with chromosomally-integrated mcherry as a model for stx2, we measure mCherry fluorescence to quantify the effectiveness and extent of our gene knockout system. Progenie can not only reduce STEC-related food recalls, illness, and waste, but can generalize to disable virulence factors or antibiotic resistance in a host of other microbes.
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TecCEM
Title
Disrupting Disruptors

Abstract
We designed a biosensor for the detection of Endocrine Disruptive Compounds (EDCs) in samples of bottled water. This biosensor works through the immobilization of protein receptor hER alpha on a piezoelectric sensor. The protein captures these chemicals, while the piezoelectric sensor senses the change of mass through a change of the natural resonance of the quartz. This signal is recieved and interpreted by a circuit of our own design and will give us information about the concentration of EDCs on the sample of water. We focused on the enzymatic degradation of these compounds as well, through the expression of a Laccase. We plan on introducing this step as part of the purification process of fresh water, guaranteeing EDC-free water to our community and reducing the risk of developing any serious illness.
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FDR-HB Peru
Title
Chococadmium: Tackling cadmium contamination in the Peruvian cacao industry

Abstract
The Peruvian cacao industry has been heavily impacted by the new European Union regulations which placed a limit of 0.8mg/kg of cadmium (Cd) in cacao products due to the carcinogenic and toxic effects on organ systems. Only 5% of Peruvian cacao currently meets these requirements leading to over 50,000 Peruvian cacao farmers reporting income loss. Our team, in collaboration with Alianza Cacao (an organization representing Peruvian cacao farmers), is working on developing a Cd detection and bioremediation system to be implemented during the cacao liquor stage of production. This way, we hope to provide a system that will impede Cd from transferring to the final products of chocolate and cacao powder.
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Nantes
Title
UlValorization

Abstract
Our project aims to reduce, and maybe even solve, a local environmental, health, and economic issue: Ulva spp. Green tides in Brittany, caused by anthropogenic eutrophication. Eutrophication induces anoxic zones that enhance the proliferation of Sulfacto Reductor Bacterias (SRB). SRB uses Ulva algae to produce hydrogen sulfide, which is deadly for a wide variety of organisms, even in small concentrations. Consequently, the whole underwater ecosystem could be damaged. Using chemical, biochemical and biological tools, our team wants to accelerate and valorize the degradation of Ulva spp., by recycling the sulfate and finding utilization for the remaining biomass (biomethane extraction, bioplastic, and compost manufacture). As it is slowly becoming a global problem, we want to raise awareness of this phenomenon through meetings, games, and social media. Thanks to collaborations with several companies that made our project possible, we expect to encourage other countries to follow up and value their green tides.
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CHINA-FAFU
Title
Microscopic Algae,“Macroscopic” Energy.

Abstract
Microalgae are among the most efficient carbon sequestering organisms known, and the CHINA-FAFU iGEM team has used synthetic biology and molecular biology as the core tools to explore the association between key genes of ascorbic acid metabolic pathway and carbon dioxide fixation and lipid anabolism in microalgae, and to construct various engineered algal strains to verify the functions of related genetic elements. At the same time, we will use mathematical models to predict the possibility of practical production applications of the algae strains. Our project can provide a reference for the design and cultivation of excellent engineered strains of microalgae, and promote the reduction of biodiesel production cost and the international trend of new energy and carbon neutral economy.
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ASIJ Tokyo
Title
ABrCaDaBra

Abstract
With 2.3 million women diagnosed and 685,000 deaths in 2020, breast cancer has become a major problem in the global community. However, early detection can help increase the survival of patients. Current detection methods include mammograms and MRIs, which can be uncomfortable and expensive, raising the need for a better alternative. For our two-year project, we plan on creating a convenient breast cancer testing kit that would be similar in design to a pregnancy test through the detection of biomarkers found in sweat and tears. This year, we focused on verifying the interaction between aptamers and biomarkers using ELISA before developing a colorimetric aptamer-based detection assay with gold nanoparticles, which we hope will translate into our testing kit next year. Through our detection kit, we hope to raise early screening rates, allowing women to go through a pain-free and simple process that is reliable and accurate to detect breast cancer.
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Sydney Australia
Title
Free Coli: Improving accessibility to synthetic biology by engineering a naturally transformable Escherichia coli

Abstract
Free Coli aimed to improve accessibility to synthetic biology by designing a genetically modified naturally transformable strain of Escherichia coli to produce a more affordable and efficient host organism that does not require chemical treatment or electroporation to become competent. Literature on natural transformation in E. coli and related bacterium was reviewed and experts were consulted to identify twenty-five putative natural transformation genes in A. baylyi for insertion into E. coli via a novel recombineering strategy for insertion of multiple gene clusters. K-means clustering used existing data on transcriptome concentration and promoter strength to model the optimal clustering of genes into eight <5kB DNA fragments. Bioinformatics analysis was conducted to assemble the genes into fragments with salicylate promoters and selectable markers. The wider community was engaged and consulted to affirm the project's purpose and align its future implementation with the UN's Sustainable Development Goal to ensure quality education for all.
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Kyoto
Title
FLOWEREVER -A colorful palette of approaches for sustaining flower life-

Abstract
Since time immemorial, flowers have been providing us with comfort and beauty on occasions ranging from mourning to celebration. While cut flowers are in high demand, they are ultimately treated as disposable products leading to large amounts of pre- and post-consumer waste. To make this intimate and essential relationship sustainable, iGEM Kyoto proposes a colorful palette of measures for each process from flower production to consumption. Our discussions with stakeholders revealed that many flowers are lost to insects or diseases. Accordingly, we attempted to kill pests with RNAi, and developed a virus detection device based on RT-LAMP and CRISPR-Cas12a. Alternative to chemicals, we suggest dsRNA and peptides to delay flower wilting. Notably, for more efficient syntheses of these eco-friendly materials, we proposed a system using asymmetric plasmid partitioning. Although fragility and mortality may add to a flower's beauty, an extended life span will allow them to be enjoyed by many.
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HK CPU-WFN-WYY
Title
Project SSS

Abstract
Inspired by Fast Fashion, which is a threat to the environment at disposal, we came up with the idea of utilizing biodegradable ‘silk'. In this project, E. coli BL21 is aimed to be engineered to produce a protein named spidroin, which is the basic unit of spider silk. Masp1 and Masp2 from dragline, flagelliform silk protein from capture spiral silk and pyriform silk protein (PiSp) from piriform silk are planned to be manufactured. In the future, the mechanical properties of silk products, like strength, will be adjusted by controlling the proportion of spidroin with different toughness and elasticity. Thus, we hope to obtain spidroin with different properties to cater for different needs, like making clothes or fishing line, hence bringing an environmentally friendly solution to Fast Fashion disposal.
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UCAS-China
Title
DeCaffi: Personalized Caffeine Intake Management Scheme Based on Synthetic Biology

Abstract
With the popularity of caffeinated beverages, more and more people are troubled by caffeine sensitivity. People drink coffee during the daytime, but toss and turn at night because of the exciting effect of caffeine. In order to help humans better manage caffeine intake, we designed DeCaffi Project. Utilizing the conversion from caffeine to theacrine catalyzed by Cdh and CkTcS, we realized caffeine removal from customers' coffee. In view of the special liquid environment of hot coffee, we improved thermal stability of these enzymes through directed evolution and software PROSS. Using immobilized enzyme technology, we constructed a cell-free system that can quickly remove caffeine in our specially designed coffee cup, in which the sensor can monitor the changes of caffeine in real time. Additionally, considering convenience and user-friendliness, our software group has designed an App, which can collect data from coffee cups and bracelets, providing users with real-time caffeine intake suggestions.
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UESTC-China
Title
Deink the dark

Abstract
Nowadays the recycling rate of office waste paper (OWP) in China is dramatically low. We find that cellulase, xylanase, lipase, and laccase will allow bio-deinking to occur, by helping the ink detach from the waste paper and decomposing hazardous substances. We also design a new surface assembly of a functional cellulosome by using Pichia pastoris. Otherwise, to bring our idea to the real world, we design a paper recycle machine (Deinker) ,which includes the transmission part, enzyme liquid smearing part, deinking part, and paper drying part. Through this device, the OWP will be reborn. In addition, 5 mathematical models are built for improving experiments, hardware, and implementation. Moreover, Human Practice has been constantly advancing as the project progresses, we design related activities from four aspects: practical issues, technology, market and industry, policy, and law to understand the exact needs of stakeholders.
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RDFZ-CHINA
Title
Killer-X

Abstract
The bacteria Xanthomonas has caused agricultural diseases worldwide. Meanwhile, the use of traditional pesticides and antibiotics is causing problems including antimicrobial resistance and social ethical issues. Therefore, we seek to find out an alternative way to treat microbial plant diseases.Through research, we found that phage therapy can be useful towards infectious diseases. However, storage difficulties and limited functional time result in the phage therapy's low popularity. Due to these problems, we intend to use synthetic biology technologies and phage delivery bacteria to develop a more efficient phage therapy towards Xanthomonas diseases.
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BIT
Title
Early screening of colorectal cancer(CRC) based on CRISPR/Cas12-coupled SlipChip for detection of miRNA

Abstract
Early screening and treatment of colorectal cancer(CRC) has been proved to be conducive to reducing the mortality rate of CRC. Currently, most diagnostic methods do not meet the requirements of rapid, sensitive and specific screening of CRC. Based on the selection of miRNA as our biomarker for its high sensitivity and specificity,we developed an ultrasensitive approach for miRNA detection based on a combination of Loop-mediated isothermal amplification(LAMP), CRISPR-Cas12a trans-cleavage and SlipChip. We also designed temperature-control and fluorescence detection modules to erect an interactive IVD system for POCT detection. Our system has been tested to be simple to operate, for completing the process within 1h,and its sensitivity reaches to fM level. Furthermore, our system has the capacity to detect different miRNAs, giving the potential to build standardized cancer detection platforms.
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YiYe-China
Title
TFPI2 Methylation in Stool Samples for Early Detection of Colorectal Cancer by Toehold Switch

Abstract
Colorectal cancer is one of the most common cancers in the world. The current method for early diagnosis of colorectal cancer remains as a big limitation. Previous reporters identified syndecan-2 (SDC2) and tissue factor pathway inhibitor 2 (TFPI2) were aberrantly methylated in Colorectal Cancer. Quantitative methylation-specific PCR (qMSP) for detecting methylated SDC2 and TFPI2 has been reported as an effective early screening method for colorectal cancer. Toehold switches are a class of RNAs with a hairpin loop that can be unfolded upon binding a trigger RNA for translation initiation of the reporter protein, which plays an important role in clinical diagnoses. In this study, we constructed a toehold plasmid and incubated it in the cell free protein expression system with methylated TFPI2 with a trigger sequence, which offers a non-invasive and visible approach for early detection of colorectal cancer in the future.
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Tongji China
Title
LOOK! Little Odor Killer

Abstract
Since the implementation of household garbage sorting regulations in Shanghai, the amount of food waste has reached a peak. However, problems caused by odor like unpleasant smell and health problems, have aroused great attention everywhere. This year, Tongji China launches the "LOOK!" project to solve the problem. We construct two kinds of bioengineered E.coli to absorb hydrogen sulfide and ammonia, which are the two main ingredients in the odor. One uses enzymes related to sulfide oxidazation(Sqr, Sdo, AprBA and Sat), converting hydrogen sulfide to sulfate, while the other uses enzymes AMO, HAO and NOD to convert ammonia to nitrate. Besides, a three-gear adjustable kill switch based on the concentration of H2S and NH3 is added to ensure biosafety. Also, we optimize our pathway by high throughput screening and machine learning. With such efforts, we hope to solve the problems the odor brought to people and downstream industries.
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Korea HS
Title
Designing a Cancer-Specific Cell-Penetrating Peptide for the Efficient Delivery of siRNA into Cancer Cells

Abstract
siRNA is a potent therapeutic agent because of its specificity and efficiency suppressing genes overexpressed during pathogenesis. However, due to lack of cell type-specific and safe delivery methods, it shows limited success in cancer treatment. Therefore, we aim to design CPP which forms a siRNA/CPP complex targeting neoplasia. Two engineered CPP based on buforin IIB - MV1 (-RLLR) and MV2 (-RLLRRLLR) - were tested. Among three types of CPPs, MV1 displayed highest specificity for A549 (cancerous). Next, our goal was to enhance the anticancer efficacy of doxorubicin (chemotherapy drug) mediated by MV1/siRNA complex. Since CYP1A1 is upregulated in doxorubicin-resistant cancer cells, siRNA was designed to target CYP1A1. Our result showed that MV1/siRNA complex efficiently suppressed CYP1A1 gene expression. Finally, MV1-DOX/siRNA complex increased anticancer efficacy of doxorubicin in A549, while showing minimal cell death on MRC5. Overall, results suggest MV1 could deliver antineoplastics and siRNA to improve anticancer efficacy.
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CKWA-China
Title
LonEase

Abstract
Cancer recurrence after surgical treatment is currently a major problem facing humans, and cancer recurrence is a huge threat to cancer patients. For example, bladder cancer has a recurrence of up to 24%-84% of patients within 5 years after surgical resection, and the overexpression of C-MYC protein is approximately related to 44%-69% of the cases. We investigated the urinary tract pathogenic Escherichia coli (UPEC) Lon protein and find it can effectively degrade C-MYC protein, thereby inhibiting the proliferation of cancer cells. We use E. coli to produce highly active Lon protease this year, and propose this protein can be designed into drugs to improve the surgical prognosis of bladder cancer populations with C-MYC gene overexpression.
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Duesseldorf
Title
Development of an in-field quick test for the differentiation of plant pathogens

Abstract
This year, iGEM Düsseldorf sets the focus on pathogen detection and differentiation. The aim is to design an adaptable quick-test with three potential applications. Firstly, to detect common plant diseases on young plants, secondly, to detect pathogens in-field before signs of infection are even visible, with the help of multispectral imaging cameras that identify plants with reduced photosynthetic activity. Thirdly, to be able to differentiate between known diseases that are hard to identify with the naked eye. The plants can be tested with our lateral flow assay consisting of two aptamers that have been carefully selected through the SELEX process. These aptamers bind specifically to either a protein expressed during the plant's stress response or the pathogenic cell itself, precisely detecting the cause of stress. To visualize general plant stresses immediately, a reporter plant was designed, whose colour changes depending on the kind of stress it's exposed to.
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British Columbia
Title
DetecTME: Untangling the immuno-modulated tumour microenvironment using a tumour-colonizing bacterial vector and reporter system

Abstract
Immunotherapies are cancer treatments which activate the immune system against the tumour. While very effective in some patients, many do not respond to immunotherapy. To personalize treatment, tissue biopsies are used to profile tumour immune activity and inform likelihood of immunotherapy response but are invasive and prone to sampling bias. Salmonella bacteria naturally colonize tumours, and have been proven safe as a therapeutic in clinical trials. Our project creates an in vivo Salmonella-based biosensor for minimally invasive diagnosis and real-time monitoring of tumour immune activity. We construct a functional screening system to discover promoters regulated by tumour immune markers across the Salmonella genome, and implement an AND-gate with a split-operon technique to detect multiple biomarkers. The biosensor will be adaptable to detect other clinically relevant biomarkers, and to deliver therapeutics. This project is a gateway to a multifunctional platform to support the growth of immunotherapy and personalized medicine
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IvyMaker-China
Title
The PET-buster Magic Yeast

Abstract
Our goal is to develop a whole-cell biocatalyst by displaying PETase and MHETase on the surface of yeast cell (Candida tropicalis) to degrade PET waste. Firstly, we chose a robust Candida tropicalis as the host and deleted the URA3 gene of genome to block the uracil synthesis by CRISPR-Cas9. In order to display the protein more efficiently, we must obtain the appropriate anchor protein. Therefore, we built a model to predict the anchor protein candidates. We used an enhanced green fluorescent protein (eGFP) as the reporter and inserted the codon-optimized GFP gene to the genome of Candida tropicalis by homologous recombination. Finally, we selected the best one from 89 anchor proteins, fused it with PETase, and then incubated the yeast with PET waste. We hope the yeast with surfaced PETase can degrade the PET more efficiently and achieved the recycling of PET waste.
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NCHU Taichung
Title
Artificial endophyte for future agriculture

Abstract
Our main goal is to use the synthetic biological strategy for achieving an example of circular agriculture.Rice could feed half of the global population, and tons of rice straws are produced after harvest. Inappropriate rice straws management, straws burning, causes problems releasing CO2, PAHs, and PM2.5.In this project, we constructed the Clostridial cellulase genes within Bacillus subtilis WB800 and the cellulosomal enzymes could be formed. It will be used for effective straws decomposition and used as a medium for culturing our beneficial endophyte.Furthermore, we constructed an artificial endophyte through versatile biostimulant, pyrroloquinoline quinone (PQQ) producing Bacillus subtilis and cultured by saccharified straw liquid.The PQQ containing broth will exogenously apply to the rice and/or other plants to promote their growth. Our example for circular agriculture could be completed by the above strategy, while agricultural waste can be recycled into valuable products and also could reduce CO2 emission.
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Brno Czech Republic
Title
Phoscage

Abstract
Our team created a system for phosphate removal from water. The system is based on modified Bacillus subtilis. This bacterium is designed to be able to measure the amount of phosphate in water. If the phosphate exceeds a certain threshold concentration, the bacterium will start to form a system to pick it up. This consists of an enzyme that will bind the phosphate into long chains (polyphosphate kinase) and protein vesicles (bacterial microcompartments, BMCs) that will enclose the phosphate chains.According to the design, these modified bacteria will be placed in water in a system that prevents their escape into the environment. After a period of time, the bacteria with the accumulated phosphates will be removed from the water, inactivated, and can be further utilized. We envision their use either as fertilizer or as a source of phoshporus for the chemical industry.
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XJTU-China
Title
Tryptophan iDream

Abstract
In our project, in order to release the pain of people suffering from mental health problems like depression,anxiety and insomnia, we apply the methods of synthetic biology on the functional change of Escherichia coli. According to our ideas, a gene circuit based on Toogle-Switch is designed to reach the goal of switching between two statuses: tryptophan-producing and proliferating, by reacting to signals of IPTG and temperature. On top of that, considering the situation of COVID-19 pandemic and lockdown, we give our project further application by constructing a hardware to help people release their pain in their home by drinking a cup of milk with additional tryptophan in it. We believe this design will actually aid people in dealing with their mental health problems, especially during the pandemic, in a much safer and user-friendly way.
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Cornell
Title
Engineering Prokaryotic Collagen-Based Scaffolds to Guide Cell Growth in Damaged Joint Tissues

Abstract
Numerous cell types must grow in specific patterns to form functional knee and elbow joints, and injury healing in these areas is difficult as these cells must all regenerate in very exact locations. This project aims to improve tissue healing by developing a prokaryotic collagen-based scaffold; this acts as a pre-programmed framework that precisely directs the growth arrangements of mouse myoblasts and fibroblasts. Strains of Escherichia coli were engineered to produce Scl2 proteins with binding motifs for integrin and fibronectin; these customizable protein products were incorporated into a hydrogel, providing a physical structure to which the mouse cells attach. The hydrogels feature optimal spatial distributions of binding motifs tailored to each cell line, causing them to grow in desired patterns due to the differences in their respective ECM affinities for integrin and fibronectin binding sites. The effectiveness of these hydrogels was characterized through cell cultures and microscopic imaging.
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CityU HK
Title
Synthetic Biology of Natural Products: Genomics-Driven Derivatization of Variecolin.

Abstract
Despite recent advances in medical science, humans are still facing some global health challenges, including emerging infectious diseases and debilitating diseases such as cancer. Hence, there is an unmet clinical need for new and effective drugs. However, a long-existing problem in drug discovery is the cost-effectiveness issue in finding effective drugs for clinical use. To address this, our team aims to develop a simple and cost-effective fungal synthetic biology platform to produce various derivatives of variecolin, a fungal natural product that shows potent therapeutic and antimicrobial properties. The platform consists of the genetically modified Aspergillus oryzae host with (1) a “terpene synthase” biobrick for the backbone synthesis and (2) a series of different “cytochrome P450 monooxygenase” biobricks to derivatize the variecolin scaffold. We have successfully obtained three novel variecolin analogues as confirmed by LC-MS and NMR analyses, and experiments are currently underway to characterize their biological activities.
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NTHU Taiwan
Title
Compine: A Novel Way to Solve Pine Wilt Disease

Abstract
Pine wilt disease is caused by specific nematodes—Bursaphelenchus xylophilus. Our goal is to generate a kind of pine tree that can synthesize siRNA that encodes important gene information to B.xylophilus, such as targeting genes related to reproduction and pathogenicity. As soon as the nematodes uptake plant cells, siRNA generate by pine trees will knock down genes that significantly influent the survival of nematodes. We build a model to simulate how the diffusion of siRNA in B.xylophilus. And the birth and dead model we build is to simulate the activity of nematodes on plant callus. We have published an article on PanSci and meet related environmental groups. Our podcast program has launched on several platforms and we designed a board game--Adaptor, introducing this environmental issue and synthetic biology to the public. We hope to generate an efficient method to solve PWD comparing to traditional methods.
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Sorbonne U Paris
Title
Chlamy'n Space

Abstract
Acute or chronic exposure to ionizing radiation (UVC, gamma rays, X rays) leads to the formation of reactive oxygen species (ROS) which affect the genome and the proteome of cells. Thus, although photosynthetic microorganisms such as Chlamydomonas reinhardtii constitute one of the main hopes for developing Bioregenerative Life-Support System during long-term space travel, their use is called into question by the decrease of efficiency of the photochemistry and by the growth arrest caused by ROS. Our project aims to make Chlamydomonas reinhardtii produce a peptide complexing with the Mn2+ ion inspired by a metabolite found in the radioresistant organism Deinococcus radiodurans and acting as an antixoidant. This study requires to demonstrate a decrease in ROS within the cell during production of the peptide and verifying the growth of microalgae cultivated in minimum medium (photosynthesis dependent growth).
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KUAS Korea
Title
Probiotic Therapeutics to Rescue Frogs from the Deadly Chytrid Fungus

Abstract
Amphibians are facing an uncertain future. A third of global amphibian species have become endangered due to habitat loss, climate change, and the lethal chytrid fungus. The invasive chytrid fungus has been prevalent in many parts of the globe but a reliable solution is not available yet. This year, our focus is on the biological control of the chytrid fungus by engineering skin symbiotic bacteria. We designed a genetic AND-gate that produces the antifungal agents (e.g. violacein) only working under sunlight warming frogs and the detection of chytrid. To avoid environmental risks, we also designed a quorum sensing kill switch based on the MazEF toxin-antitoxin module. We surveyed the skin microbiome of frogs for a potential chassis of our design. Our therapeutic approach will contribute to protecting susceptible amphibian species from the invasive chytrid disease.
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SZU-China
Title
Gutail Floractory - A customized platform for IBD cocktail adjuvants based on intestinal engineered bacteria

Abstract
IBD, as a chronic intestinal inflammatory reaction, recurs frequently and is a potential cancer risk. The disease is currently incurable and exerts many different stresses on patients. This year, we aim to develop a 'cocktail' of customized, genetically engineered intestinal probiotics as an adjuvant to alleviate the recurrence of IBD and improve patient's quality of life. Based on E. coli Nissle 1917 and L. lactis, we express four related substances to target IBD: butyric acid, which can consolidate the intestinal barrier and inhibit inflammatory pathways; human antimicrobial peptide LL37 for LPS resistance; SOD to reverse peroxide stress state; and BSH to reverse abnormal bile acid metabolism and reduce inflammation. To successfully implement our project in the context of potential patient needs, we write a business plan and design a series of hardware devices. We believe our project will be practical and can offer far-reaching benefits to many future patients.
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Patras
Title
PGasus: A comprehensive workflow for the identification & functional characterization of PGx variants

Abstract
iGEM Patras 2021 goal is to characterize the impact of genomic variants, derived from targeted panel-based resequencing using a portable next-generation sequencer, on the enzymatic activity of CYP2D6 and CYP2C19 enzymes, responsible for the metabolism of clinically used drugs, using in vitro assays. These variants will be engineered into gene constructs, via site-directed mutagenesis in eukaryotic 293FT cells, encoding these enzymes and will be subsequently characterized as deleterious or benign, and compared against in silico prediction scores. Furthermore, to validate the mechanisms underlying the molecular interaction between the mutated CYP isoforms and the chemical substrates, we used molecular simulation methods like docking, to confirm the potential deleterious effect of the identified PGx variants.
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Greece United
Title
Epione: One Less Painful Step

Abstract
Osteoarthritis is the most common musculoskeletal disease around the world, often leading to chronic pain and severe paralysis. Osteoarthritis is characterized by an intense inflammation cycle, in which chondrocytes produce certain proteins that cause cartilage degradation. Our approach aims to halt this inflammatory cycle by inserting specifically transfected cells in the osteoarthritic joint. The cells produce synthetically designed exosomes that contain miRNA-140. miRNA-140 has been shown to inhibit the expression of certain cartilage destructive proteins and promote the expression of regenerative agents. The exosomes are also designed to carry a Chondrocyte Affinity Peptide (CAP) in their membrane which guides them to deliver their cargo specifically towards chondrocytes. This project includes in-silico and in-vitro experiments that study the efficiency of the proposed plasmid to produce exosomes in HEK293T cells. Project dissemination activities include a novel web application to communicate osteoarthritis risks to the public.
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DTU-Denmark
Title
Engineering K. Phaffii for methane utilisation

Abstract
Methane is a potent greenhouse gas contributing to climate change. The UN Sustainable Development Goals clearly emphasize the immediate need to combat climate changes to promote sustainability. Our project, Pheast, endeavored to design and test an engineered cell factory in yeast to use methane as a feedstock. We used Komagataella phaffii (P. pastoris), a methylotrophic yeast, to engineer this cell-factory since it has been used extensively in industrial protein production due to its high protein-secretion as well as ease of protein purification. To make the cell factory effectively use methane as a carbon source, recombinant pMMO subunits were incorporated from a natural methanotrophic bacteria, M. capsulatus. This way, methane is converted to methanol and K. phaffii can further use its native metabolic pathway to produce protein of interest. In the future, we believe this project will open the arena to produce many proteins of interest using this engineered, methane-consuming cell-factory.
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GA State SW Jiaotong
Title
Heterogeneous Expression of Recombinant Gas6 and EGF Proteins for Curing Alopecia

Abstract
In research published in the past year, two critical proteins, Gas6 and EGF, have shown promise for treating stress-related and androgenetic alopecia. Plasmids were constructed for transformation into Pichia pastoris, E.coli, and Bacillus subtilis respectively. Gas6 proteins will be purified from Pichia pastoris and EGF proteins will be expressed in E.coli using a T7 promoter to produce a cell-free protein treatment for hair loss. As a possible alternative, two plasmids with the coding sequence for each protein and a suicide switch (to prevent unwanted growth of the bacteria after treatment) were designed to create a living-cell protein expression system in B.subtilis which could be applied to the microbiome of the scalp.
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Waterloo
Title
NeuroDetech

Abstract
Attention-Deficit/Hyperactivity Disorder (ADHD) is a neurodevelopmental disorder characterized by inattentiveness, an inability to focus, and restlessness. Current ADHD diagnosis procedures require a series of psychological assessments that are qualitative in nature and subject to potential clinician bias. Waterloo iGEM's 2021 project, NeuroDetech, is a quantitative tool that aids in the diagnosis of ADHD and reduces the margin of bias. NeuroDetech consists of a series of microfluidic capillary flow assay lab chips designed using process engineering methods. Each lab chip contains a binding molecule that detects an ADHD-associated biomarker or gene marker in urine. Through protein engineering and computational protein redesign methods, the binding molecule can be adapted for the sensitive detection of a variety of ADHD-associated markers. Each lab chip and optical detector emits a signal over time that is visualized on a software app, providing healthcare professionals with insight into an individual's risk for ADHD.
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DNHS SanDiego CA
Title
Vigilantly Optimizing Cancer Detection: Using E. coli to detect volatile organic compounds in the breath

Abstract
Lung cancer remains the deadliest cancer type, and is often undetected until stages 3 or 4, when it becomes difficult to treat [1]. W decided to investigate a noninvasive method: VOC (volatile organic compound) detection using a breath test. Our goal is to use the K-12 strain of E. coli bacteria that will produce a quantifiable color change in response to the presence of a VOC. Using published research, we identified a set of VOCs whose concentration in the breath is higher in lung cancer patients: propanol, trimethylbenzene, toluene, ethylbenzene, styrene, hexanal, acetone, and isoprene [2]. We will insert promoters that are induced by these VOCs into plasmids and a GFP gene that will produce a color change, transforming E. coli with these plasmids. This product would be implemented as a test that could use one breath sample to detect the levels of 8 separate VOCs.1. https://www.cancer.org/cancer/lung-cancer/detection-diagnosis-staging/how-diagnosed.html 2. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6044508/
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Moscow
Title
miPression - the diagnostic test system for depression detection based on CRISPR/Cas.

Abstract
We are the Moscow team, and the main goal of our project - miPression - is to design a test system that can detect depression and distinguish it from other psychiatric diseases. Our diagnostic test will use miRNAs circulating in blood as biomarkers. We are going to measure it by two-step technology: first, rolling cycle amplification (RCA); second, the connection of two dСas9, which are linked with the half of GFP, to the product of the RCA. The concentration of our target miRNA will be defined based on the brightness of fluorescence. As a result, our project will help to solve the problem of misdiagnosed and undiagnosed cases of depression. In the long term, our development may help to reduce public stigmatization and break down false stereotypes about mental disorders.
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Hamburg
Title
E.T. - The ElectronTransferrestrial

Abstract
Terpenoids are a large class of plant secondary metabolites used as food-flavors, fragrances or pharmaceuticals like artemisinic acid. Today, terpenoids are either extracted from plants they naturally occur in or produced synthetically using petrochemical starting materials.Only in a few cases have biotechnological approaches been successfully implemented for the production of terpenoids. Despite their chemical diversity, mainly two enzyme classes are required for terpenoid biosynthesis: cyclases and oxygenases. While cyclases are widely used in prokaryotes, oxygenases naturally occur as membrane-bound protein complexes. For functional expression in E. coli, soluble versions as well as compatible reductases are required. Additionally, protein expression places significant strain on the energy metabolism of E. coli.We are addressing these problems by creating fusion constructs of oxygenase and reductase proteins, as well as increasing the availability of NADPH. Advances in this field will lead to more sustainable and economical terpenoid production in the future.
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Tongji Software
Title
Phage-MAP

Abstract
Phage therapy has become one of the most promising alternatives to antibiotics in the treatment of bacterial diseases, and identifying phage-host interactions helps to understand the possible mechanism through which a phage infects bacteria to guide the development of phage therapy. Compared with wet experiments, computational methods of identifying phage-host interactions can reduce costs and save time and are more effective and economic. Through two methods, alignment-based and alignment-free, our team attempted to find accurate Phage-bacteria interactions, contributing to phage therapy, and further contributing to the application of phage as a model organism in synthetic biology.
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IISER Bhopal
Title
BLAST : Bifidobacterium Longum induced Apoptosis using Smac and Trail (Therapeutic for Solid tumor cancers)

Abstract
Cancer, the second leading cause of death in the world, registered 19 million cases and about 10 million deaths in 2020 globally. Cancer treatment is both financially and mentally traumatizing for patients as current therapies suffer from a plethora of side effects and are very expensive. Our project presents an innovative approach to treat cancer using apoptosis-inducing peptides SMAC & TRAIL. TRAIL binds to the cancer cells extrinsically, thereby activating caspases, whereas SMAC induces apoptosis intrinsically and also sensitizes TRAIL-resistant cancer cells. Our chassis organism, B.longum, is an obligatory anaerobe and grows only in hypoxic regions providing specificity to target solid tumours. Furthermore, it has a lactate-based KILL SWITCH to ensure it gets killed outside the cancer environment and an AND GATE ensures secretion specificity to cancer cells only. We'd be establishing the Proof of Concept in E.coli. The objective is to design cheap and viable therapy for cancer treatment.
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Baltimore BioCrew
Title
Joy-Biotic: Challenging Mental Illness & Its Stigma One Flavored Drop at a Time

Abstract
The COVID-19 Pandemic has brought an increase in mental illness rates. The group most critically affected by this increase is high school students, the peer group of The Baltimore BioCrew. To challenge the stigma and the increasing rates of mental illness, the BioCrew has created Joy-Biotic, a flavored water probiotic that will increase serotonin production by producing 5-HTP, GABA, and acetylcholine in Lactobacillus and E. coli Nissle. These molecules increase neurotransmitters in the brain. To prevent serotonin syndrome, we engineered a negative feedback loop that would regulate the levels of serotonin. To raise awareness about mental health stigma, we have created a podcast interviewing mental health professionals, a social media campaign, an educational comic, and a course teaching high school students about the relationship between the gut biome and mental health. Through oral ingestion of Joy-Biotic, the BioCrew plans to fight mental illness one flavored drop at a time
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BIT-China
Title
Creative Tasting Officer

Abstract
Taste is the soul of food. Food companies all over the world are looking for better ways to quantify the basic flavors of newly developed food products. In the food industry, human sensory panel and electronic tongue are most commonly used to evaluate taste. However, human sensory is costly and subjectively affected, while one electronic tongue cannot measure the taste of multiple food. This year BIT-China devotes to design a Creative Tasting Officer-a yeast cell biosensor to quantify the basic tastes of umami, sweetness and bitterness. Genetic circuits are designed to express human taste receptors heterologously in Saccharomyces cerevisiae, and the intensity of fluorescences would quantitively reflect the respective taste degree of food samples. Furthermore, we develop a series of Soybean based Creative Seasonings and a Flavor Card Website to help people discover their favorite taste. Everyone can click the link and find the satisfying flavor with personalized color panel!!!
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CPU CHINA
Title
Construction of MnP-AAO-HFB1 composite system based on CRISPR/dCas9 programmable assembly technology for polyethylene plastic oxidation.

Abstract
Examplified by polyethylene, undegradable plastics have a degradation period of hundreds of years under natural conditions, causing severe environmental crisis worldwide. Manganese peroxidase (MnP) uses hydrogen peroxide (H2O2) to produce high-redox-potential Mn3+, possessing a PE-degrading potential. Our project takes MnP as our key PE-degrading enzyme, employs aryl alcohol oxidase (AAO) to provide H2O2 for MnP, and utilizes hydrophobin-1 (HFB1) to increase substrate accessibility, as well as combines SpyCatcher-SpyTag system with CRISPR/dCas9 system to anchor MnP, AAO and HFB1 onto one double-stranded DNA scaffold according to certain spatial order, distance and proportion. The final complex works as a molecular machine that can adhere and degrade PE in a green and swift way
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UMA MALAGA
Title
Oil Together Painting, art from waste

Abstract
How about converting a residual source into a source of creativity and expression? For every single litre of waste cooking oil (WCO) spilled, more than 1,000 litres of water are polluted. While classic recycling is unattractive, synthetic biology is a powerful tool in order to change this view and attract people to the new paradigm of a circular bioeconomy. To do so, we aim to expand the molecular toolbox of the yeast Yarrowia lipolytica for developing a bioprocess that creates value from waste. This microorganism stands out due to its ability to use oleaginous substrates to grow. First, we are implementing the full carotenoid biosynthetic pathway in this chassis. Besides, we are optimizing the stock of WCO as its carbon and energy source to produce sustainable paints based on these carotenoids pigments. Finally, we will improve the extraction of pigments and preparation of bio-paints for scaling-up.
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BOKU-Vienna
Title
Friendzyme - a novel therapeutic platform in case of Irritable Bowel syndrome, and related disorders

Abstract
About 15% of the worlds population suffer from bowel disorders like irritable bowel syndrome (IBS), and related disorders. These are triggered by so-called FODMAPs (fermentable oligo-, di-, mono-saccharides and polyols), which are non-digestible food components that lead to bloating, abdominal pain and other symptoms. So far the only therapy is a highly restrictive diet. We want to change that by addressing neglected gastrointestinal diseases and tackling one of the most common FODMAPs: Fructans, a fructose polymer. Our goal is to develop a novel therapeutic platform by the use of lactobacilli that secrete fructan-degrading enzymes. We encapsulate them in a biocompatible scaffold that keeps them safely inside while allowing the release of the produced enzymes. The surface of the scaffold is designed to have muco-adhesive properties which enable site-targeted enzyme delivery over a prolonged timespan. With this concept we want to take oral enzyme therapy to the next level.
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CTR Alberta Canada
Title
Detecting oxybenzone via production of LacZ gene induced by GEM protein in engineered S.cerevisiae

Abstract
Oxybenzone is a chemical compound found in sunscreens that serves as an active ingredient to block ultraviolet (UV) rays. Oxybenzone contributes to coral bleaching by accumulating in coral tissues and reducing sunlight availability for photosynthetic organisms due to its UV-absorbing properties. Through the genetic engineering of Saccharomyces cerevisiae, oxybenzone concentration levels in water bodies can be detected. The estrogenic properties of oxybenzone allow it to bind to a synthetic fusion protein containing a human estrogen receptor which will result in the expression of the LacZ gene. This causes the enzyme beta-galactosidase to be produced and breakdown its substrate. This produces an electrochemical signal which can be measured using the Field-Ready Electrochemical Detector (FRED) and analyzed to determine oxybenzone concentration. This will allow for greater understanding of the issue of coral bleaching, and provide a tool to assess the impact of further steps being taken to protect these aquatic ecosystems.
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RUM-UPRM
Title
R-DetoX: A Biodegradation Solution for RDX Contamination in the Anones Lagoon in Vieques, Puerto Rico

Abstract
The Anones Lagoon in Vieques, Puerto Rico, has been impacted by the presence of xenobiotic contaminants, whose concentrations exceed established limits by the EPA, with one of concern being a recalcitrant explosive compound, 1,3,5-Trinitro-1,3,5-triazine (RDX). Our goal is to design R-DetoX, a two-component genetic circuit for the detection and biodegradation of RDX. An RDX-inducible sensor element is transcriptionally coupled with the quorum-sensing (QS) signal biosynthesis and transcriptional regulator genes, luxI and luxR, respectively. The second component is induced by acyl-homoserine lactone (AHL)mediated QS, activating PluxR,, which initiates the transcription of the xplA/B system. As a result, denitrification and ring-cleavage of RDX occurs, reducing the compound's recalcitrance. A fluorescent reporter follows the catabolic component, allowing for quantification of biodegradation gene expression through fluorometry. The biodetection/biodegradation of RDX by R-DetoX will allow recovery of the biodiversity and provide new tools for environmental monitoring and remediation.
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GreatBay United
Title
Artificial Limulus Amebocyte Lysate

Abstract
Bacterial endotoxin testing is indispensable for disease diagnosis and the launch of new drugs, demand is increasing year by year. The limulus blood reagent taken from the blood of horseshoe crab is currently recognised as the most reliable detection method, but 600,000 horseshoe crabs are dedicated to this every year. This project draws inspiration from bacterial endotoxin detection and horseshoe crab protection, and plans to develop a new, efficient and accurate method to meet the market needs of endotoxin detection. Based on the principle of the limulus hemagglutination reaction, three systems are designed to simulate the limulus reagent reaction. It is hoped that while filling the technological gap, members from GreatBay_United can saves the future of an endangered species.
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GreatBay SZ
Title
ARTAG - Artificially Revolutionized Tracking Art Gadget

Abstract
Many artworks now have serious fraud problems. These counterfeit products have significantly harmed the rights and interests of consumers and even more disrespect for artists. However, the current appraisal of artworks is poor, complicated and expensive. In this regard, GreatBay_SZ has developed ARTAG - Artificially Revolutionized Tracking Art Gadget. In ARTAG, yeast spores, which have strong tolerance and adhesion, were used as an ideal carrier for long-time DNA barcode storage. Customized DNA-encode barcode information was integrated into the genome of the yeast through homologous recombination. A CRISPR-based nucleic acid detection system was introduced to detect sequences quickly and efficiently in terms of information decoding. In general, ARTAG offers an accurate, rapid, durable, invisible and cost-efficient spore detection system to verify the authenticity of artworks. It provides new possibilities for all anti-counterfeiting technologies and markets.
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RUBochum
Title
Project Platylicious

Abstract
Livestock farming, especially dairy and meat production, contributes to a large extent to CO2 emissions and thus to climate change. More and more people are questioning their diet in order to find new ways of contributing to the protection of the climate. Our goal is to use the DNA of cows and the platypus to produce milk proteins with Pichia pastoris that do not differ in their molecular structure from those of these animals. This would offer an equivalent substitute for those people who do not necessarily want to switch to plant-based milk, but do not want to contribute and enforce global warming and animal exploitation. By producing platypus-yoghurt we try to do our part while simultaneously raising awareness for the platypus, a species threatened by dairy farming.
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Humboldt Berlin
Title
Target Taxi – Salmonella minicells as drug delivery system

Abstract
Cancer is a global threat to humankind being the cause of million deaths per year. Chemotherapy often remains the last resort, with the major drawback of being non-specific. To overcome this limitation we aim to design genetically modified minicells derived from non-pathogenic Salmonella Typhimurium. We plan to equip our chassis to specifically target cancer tissue and inject on site a cell growth-inhibiting peptide into malignant cells. In a first approach, we validate the chassis-cell interaction by modifying the minicells with lectins that specifically adhere to a targeted cell line. Subsequently, via the type-III secretion system of Salmonella, the anti-cancer peptide Pep8 will be translocated into the cancer cell, and inhibit further uncontrolled cell growth. We believe our approach will provide insights to crucial aspects of cancer research such as therapy-specificity and limitation of side effects.
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ULaval
Title
aSAP: a Solution Against maple Polymers

Abstract
Maple syrup is a Canadian emblem. It's a big part of our economy and culture. In 2021 alone, Canada produced 45M litres of maple syrup. Maple syrup is produced by boiling maple sap to make the sticky sweet syrup we Canadians know and love. Sometimes, contaminating microorganisms create exopolysaccharides in the syrup. These polymers are dextrans: very long sugar molecules that ruin the syrup's taste and texture, making it very viscous. When this occurs, it creates ropy maple syrup. This syrup cannot be sold, and so producers must pay for it to be destroyed, creating a waste of food, energy and money. We created a treatment using a designed dextranase to put into the barrels of ropy syrup to make it usable again for producers. Over the past two years, our team was able to express, purify, characterize and do the proof of concept of our dextranase.
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PuiChing Macau
Title
Turning food waste into biodegradable plastic (PLA) using engineered E.coil

Abstract
Pollutions from plastic and food waste are both pressing matters nowadays. In synthetic biology, researchers have been developing bacteria that decompose food waste and bacteria that produce bioplastic. In this project, in hope of turning food waste into bioplastics, we first engineered acid-tolerant E. coli which survive the acidic food fermentation process. Through overexpressing fabB gene, we demonstrated significant improvement in E. coli proliferation under acidic conditions. Next, we further engineered E. coli which produces bioplastics, Polylactic acid (PLA). We improved the bioplastic-producing constructs from previous researchers and iGEM team by 1) optimizing heterologous genes PhaC-PCT; 2) adding a Phasin gene to the construct. With IR spectroscopy, we proved that our E. coli can produce PLA. Using Nile red to quantify the PLA, we also demonstrated that our constructs can increase PLA production. In addition, we also built computational models and hardware to maximize PLA production from food waste.
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Thessaly
Title
Amalthea: a complete prevention toolkit to gut dysbiosis

Abstract
Diet-related non-communicable diseases (NCDs) are one of the greatest pandemics of the 21st century. Most of them are reflected on the metabolic capacity of the gut microbiome, thus making it an attractive target for further research and interventions. The impairments on the microbiome's metabolic profile, called gut dysbiosis, is characterized by altered concentrations of Short Chain Fatty Acids (SCFAs). Thus, we are developing a bioelectronic capsule that utilizes a biosensor which correlates the amount of gut SCFAs with gut microbiome functional capacity, also supported by a kinetics-oriented in silico analysis. The data is digitized and transmitted to a mobile application, where a multidisciplinary team of health experts evaluate the data received. If dysbiosis is detected, we alleviate it by the usage of a living biotherapeutic, synbiotic nutritional supplement in a personalized manner.
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ICJFLS
Title
Disposable and Biodegradable Straw Made from Amylose Produced by E.coli

Abstract
In January 2021, the Chinese government started to enforce the ban on the use of disposable plastic straws in the food industry as they cause great harm to the environment.At present, paper straws are the most popular alternative to plastic straws. However, paper straws have many defects including bad taste, easy to crack and short lifetime.Amylose is a polysaccharide chain linked by a-(1,4) glycosidic bond with D-glucose. It has poor water solubility, anti-swelling and fat insolubility, which can be used in the production of food packaging materials.In our project, we introduced two enzymes involved in amylose synthesis into E. coli: ADP glucose pyrophosphorylase containing 2 subunits ADG1 and APL1, and granule-bound starch synthase 1 (GBSS1). Once our project is completed, amylose will be synthesized and used for the production of biodegradable straw. It will replace the plastic and paper straws with a broad prospect.
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NNU-China
Title
Construction of the BL21 (DE3)-derived variant strains library to rapidly improve the antimicrobial peptides production

Abstract
With the overuse of antibiotics, more and more antibiotic-resistant bacterial strains have emerged. Antimicrobial peptides (AMPs) are considered the promising alternatives to antibiotics. Escherichia coli is the extensively used host for the production of AMPs. However, the expression of AMPs genes is low due to the toxicity of the AMPs. In order to improve the production of various AMPs, the BL21 (DE3)-derived variant strains library with different RBS sequence of T7RNAP was constructed by base editor and CRISPR-Cas9, respectively. Excitingly, the CRISPR-Cas9 system enables to obtain 87.5% of the theoretical values variants in a single editing. Furthermore, the high throughput host screening platform for AMPs production was built, and it takes only 3 days to get the best expression host for certain AMPs production. As proof-of-concept, the production of ten hard-to-express AMPs were greatly improved. We aim to protect the human health and environment by reducing the abuse of antibiotics.
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TAU Israel
Title
Communique – A generic platform for designing genes with selective activity in microbial communities

Abstract
Microbiomes include networks of microorganisms that share genetic information viahorizontal gene transfer. Most bacteria in nature are organized in such interconnectedcommunities; thus the ability to engineer microbiomes constitutes an important frontierfor the field of synthetic biology.Effective engineering of microbiomes heavily relies on selectivity: the ability to make the desiredchanges (e.g. introducing new functions) to certain species but not others. Selectivity isalso critical from a biosafety perspective as the introduction of genes into unwanted hostscan cause large ecological impacts.We have developed the first computational tool to selectively engineer microbiomes,along with experimental means to evaluate it. By addressing the main biophysicalprocesses related to gene activity and expression, our software designs microbiome-specific plasmids that are expressed optimally in certain microorganisms while havingimpaired expression in others. Our tool can be applied to any plasmid and microbiome.
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Aboa
Title
The Lac Case - Utilization of laccases for pharmaceutical waste detoxification

Abstract
Pharmaceutical waste is one of the most deleterious pollutants in the Baltic Sea. Especially the non-steroidal anti-inflammatory drug diclofenac is causing severe harm to this delicate ecosystem. The current removal efficiency of diclofenac is only 27% at our local wastewater treatment plant. The project objective was to contribute to the development of a microbial wastewater treatment system for the detoxification of this compound. The approach was to overexpress and extract three heterologous laccases, specific enzymes which are capable of catalyzing the conversion of diclofenac into less harmful derivatives, in engineered E. coli. We were able to successfully produce and purify CotA (from B. subtilis) and CueO (from E. coli), of which CotA was shown to have catalytic activity in vitro. Implementation of this work would include the expression of this laccase in photosynthetic cyanobacteria in a closed bioreactor system, integrated as a part of the wastewater purification process.
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NJMU-China
Title
Stairway to starlight:Maternal prevention and offspring treatment of autism induced by maternal infection during pregnancy

Abstract
Autism is the fastest-growing developmental disability. According to American CDC, the prevalence of autism in the U.S. has increased from 1 in 68 births in 2010 to 1 in 54 births in 2020. However, to date, there is no effective drug to treat ASD. Apart from genetic factors, the potentially dangerous environmental factors to the maternal generation, such as Maternal immune activation(MIA) and Gestational diabetes mellitus (GDM), can also be the cause of ASD. According to the gut-brain axis theory, whether it is maternal immune activation or genetic factors, the etiologies of autism are all related to the gut microbiome. Children with ASDs are also what we call ‘Children From the Star'.Our project aims at building up a stairway to the starlight, using engineering probiotics to prevent autism caused by maternal infection during pregnancy and treat children, thus structuring a way to health for the ‘Children From the Star'.
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BJ101ID
Title
Enhancing Recycled Paper

Abstract
Ever since the industrial revolution, mass manufacturing of products through assembly lines and technologies emerged. Paper, being a commonly used household and working item, had production boost from 20th century. Meanwhile, people realized how their actions damaged environment, which pushed the development of Recycled Paper. Up to now, recycled paper process is still not "environmentally-friendly" and the quality is not as good as first-hand one. BJ101ID are dedicated to solving this problem using synthetic-biology. We explored the possible effects of additives sericin and chitosan on recycled paper's strength and elasticity and proved the improvement in lab. Through E.coli, we were able to produce these two substances by genetic modification. Further experiment and modeling formulated a preeminent result with high quality and spotless paper. At the same time, we developed partnerships and collaboration with other iGEM teams and educated the new generation with what we know through seminars and online conferences.
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BUCT
Title
Delivering gaba and 5Htp through fatty acid lowering (DeliGHtFAL)

Abstract
The fast-paced life of contemporary society renders people suffer from obesity and anxiety seriously . Focusing on two grand challenges: overwork-induced obesity and anxiety, as they reflect disorders in both gastrointestinal and nervous systems, we based our approach on the current theories of the ‘brain-gut axis'. *E. coli* Nissle 1917 was used as a chassis with enhanced capacity of β-oxidation of fatty acid, thereby increasing fatty acids consumption from diets to facilitate weight loss. Meanwhile, with a synthetic biology thinking, GABA and 5-HTP with anxiety-relieving effects were synthesized from the catabolic products of fatty acids in our engineered bacteria.
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SZTA RMG Szeged
Title
Developing a novel, inexpensive method to detect small RNAs in biological samples

Abstract
Our team aims to create an entirely new way of RNA detection. This method can serve as an alternative for the expensive rtPCR method in miRNA- or other small RNA-detection in research or medicine. We collaborated with GPs and researchers.The method is based on a unique idea of RNA amplification loop. We use magnetic beads coated with single-stranded probe DNAs on which the target RNA acts as a primer for second-strand synthesis. The resulted dsDNA is the starting point of a transcription process in the same medium. The newly synthesised RNAs start the process again and again, thus producing a high amount of dsDNA detected by increasing SYBR Green fluorescence.We calibrated a simple fluorimeter and proposed a self-designed and 3D-printed adaptor for PCR tubes as inexpensive hardware for qualitative analysis. A software tool was also developed to create appropriate DNA-probe sequences for any small RNAs.
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DKU
Title
“Cyano Killer”: Biological control of cyanobacterial blooms

Abstract
Harmful algal blooms have been a severe environmental issue threatening water quality and habitat. Efforts have been made through physical and chemical methods, but with little influence. Biological methods might be more efficient and eco-friendlier. We produce and deliver algicidal polypeptides with E. coli as the agent. The polypeptide works by penetrating cyanobacteria, which triggers cell aggregation and division. The physical delivery unit is designed using mathematical modelling. It is a floating unit with adequate mobility and precise targeting that releases the drug at a controlled rate. For human practices, we asked for advice from algicidal companies and bureaus. We also made efforts in spreading environmental awareness, especially by presenting our iGEM project at the School Club Fair and publicizing our discovery through the DKU MediHealth podcast.
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UParis BME
Title
ExoSwitch – liquid biopsy for early-stage cancer screening

Abstract
Despite the growing arsenal of antitumor therapies, cancer remains the 2nd leading cause of death worldwide. A tumour starts developing asymptomatically, becoming less responsive to therapy by the time of diagnosis. We can detect cancer sooner by screening bodily fluids, such as blood or urea, for presence of tumour-associated biomarkers. These biomarkers include proteins, nucleic acids, and circulating tumour cells. However, a large-scale screening initiative requires a test for a dynamic marker that appears early during the disease and is shared by multiple tumours, such as microRNA (miRNA). Our prototype microfluidic chip, ExoSwitch, makes nucleic acid detection acessible and simple. By utilizing de novo designed RNA machines termed toehold switches, it allows rapid detection of multiple cancer-linked miRNAs. The “lab-on-chip” technology offers parallel testing for multiple targets and requires minimal equipment to perform the assay. Combining low cost and easy distribution, ExoSwitch can bring accessible tumour screening to patients worldwide.
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Austin UTexas
Title
OCTOPUS (Ocean Clean-up Targeting Oil and Plastics Using Synthetic-biology)

Abstract
Ocean pollution by crude oil and its derivative, plastic, is a pervasive environmental issue. With OCTOPUS, we intend to create a P1 phage-based system for delivering surfactant and plastic-degradation genes to marine bacteria to disperse oil slicks and degrade PET plastics, creating more accessible droplets and simple hydrocarbons to allow both pollutants to be metabolized by native hydrocarbon-degrading marine bacteria. We are focusing on the surfactant genes Ranaspumin-2 and Latherin which work individually, and the plastic-degrading genes PETase and MHETase which work together. Using Golden Gate assembly, we are creating plasmids containing either surfactant or plastic-degrading genes plus a packaging sequence that allows P1 to take up the plasmid. Once these phages have been successfully produced, we will analyze their lysates with a variety of assays such as the oil spreading assay for measuring surfactant activity and the blue-dye assay for measuring plastic degradation.
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GDSYZX
Title
Reconstruction of a highly efficient feather degrading Streptomyces sp. SCUT-3 for waste recycling

Abstract
Feather is one of the highest protein-containing resource in nature but is poorly reused. Previous reports have showed that Streptomyces sp. SCUT-3 can efficiently degrade feather. Although co-overexpression of CDO1 and protease Sep39 improved SCUT-3's feather degradtion efficiency distinctly, SCUT-Ocdo1-sep39 yield fewer cells than SCUT-3. In this study, we revealed how to use different promotesr such as pro22610, pro24880 and pro1380 to increase SCUT-3's feather degradtion efficiency without influence the growth of Streptomyces sp. SCUT-3.
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BS United China
Title
Quorum Sensing Disabler for fooD Decontamination

Abstract
Food bacterial contamination impacts human health everyday. Food is easily colonized by pathogenic microorganisms, using quorum sensing (QS) to form biofilms on food surfaces, during production, processing, transportation, and serving. QS is the regulation of gene expression reacting to cell density and is used by both Gram positive and Gram negative bacteria to regulate various physiological functions. Our synthetic biology framework consists of two E.coli constructs, Part1(E.bsuahlscout), possessing LuxR to sense and combine AHL in the environment, which will be activated and initiate the expression of mcherry report gene, and Part2(E.bsuahlterminator), containing quorum quenching acylase pvdq fused with GFP to convert AHL into LHL, blocking QS signals. To improve LL-37 peptide generated by Jilin_China, we conjunct it to our Part 2, so P. aeruginosa are exposed to it and killed, and will live under a low-AHL environment. Thus, biofilms are not formed, and food decontamination is achieved.
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Jilin China
Title
Mosquiet: A Environment Friendly And Quieter Biological Mosquitoes Trapping System

Abstract
Currently, more and more people suffer from mosquitoes and mosquito-borne disease. Although variety of methods have been developed to protect people from mosquitoes by using chemicals, they all have some defects such as slight toxicity, limited scope of action and so on. To solve this problem through biological method, a type of E.coli has been engineered to effectively attract mosquitoes in our project by releasing the three-gases-mixture ,including L-lactic acid, acetic acid and ammonia. Besides, the CRISPRi/dCas9 technique has been applied to regulate the use of carbon to increase the production of gas. Meanwhile, an acid-responsive dynamic control system and a quorum-sensing system has been added to ensure the normal growth of E. coli in a weakly acidic environment. In addition, sucrose-related suiciding system and carbon source utilization system have been introduced into the bacterium to ensure biosafety and meet the demand of household device.
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ZJUT-China
Title
CRISPR/Cas9-based Cell-Free Biosensors for RNA Biomarkers

Abstract
According to literature, various RNA molecules in urine or blood have proven to serve as biomarkers for the diagnosis and prognosis of various diseases. Herein, we are developing a cell-free RNA biosensor which comprises a cell-free system, engineered DNA transcription templates and RNA-responsive CRISPR/Cas9 System. Once the sensor is exposed to specific RNA, the visible outputs from deGFP can be visualized by naked eyes.We hope our project will provide patients with a simple, safe, low-cost diagnostic method.
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NDNF China
Title
Hidro - Say Hi to the World

Abstract
There is no denying that synbio is a hugely potential field. But for engineered strains to function beyond laboratories, some obstacles are holding them back. NDNF_China hopes to help these strains work beyond the laboratory in a safe, stable and traceable way. Here we present Hidro: a hydrogel system enclosing engineered bacterial strains. The outer layer of Hidro is a compact shell, offering both protection and containment, preventing the strains from escaping into the wild; the inner core of Hidro provides a supportive environment for them under harsh conditions, thus enabling their stable function; A genome-integrated Tracing and Control system offers tracking and specific killing of engineered strains in case of emergencies. We have experimentally demonstrated that Hidro can be implemented in diverse scenarios, such as heavy metal sensing, food-quality detection, drug secretion, etc. The Hidro system has the great potential to promote synthetic biology applications beyond the laboratory.
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BJU China
Title
Indyego: application of bio-synthetic tyrian purple on semiconstructor

Abstract
Chemical production of indigo and its derivatives will result in heavy contamination of environment. In order to improve our environment, we use genetically modified Escherichia coli to produce derivatives of indigo such as 6,6-dibromoindigo, forming indigo dyes without generating large amounts of wastewater and harmful chemicals. In addition, in order to response to the green and sustainable development, we apply 6,6-dibromoindigo on dye-sensitized solar cell to release the heavy load on semiconductor production. Also, this method gives a new way on the synthesis biology and industry engineering.
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SJTang
Title
Eco.H: Sustainable Energy via Photosynthetic Fermentation

Abstract
Compared with traditional means, biohydrogen production is cost-effective and owns unique environmental advantages. Herein, SJTang modified E. coli and R. palustris, a purple non-sulfur bacterium notable for its hydrogen production ability. In both strains, photosynthetic biohybrid systems are introduced by interfacing iodine-doped hydrothermally carbonized carbon on the cell surface, facilitating photoelectron transduction. Moreover, oxygen-tolerant [NiFe]-hydrogenase is expressed heterologously to enhance versatility and efficiency. Genes encoding for biofilm and uptake hydrogenase in R. palustris were knocked out to maximize yield. Upon modification, the co-culture of E. coli and R. palustris demonstrated a mutualistic relationship in hydrogen production by exchanging essential carbon and nitrogen sources. A low-cost bioreactor capable of fermentation, gas collection, and quantitative component analysis is developed and packed into an integrated open-source solution. Through metabolic modification and hardware innovation, SJTang hopes further to improve hydrogen production efficiency and lower unit costs.
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IISER-Tirupati India
Title
OviCloak: A Novel Contraceptive for Uterus Owners

Abstract
The modern contraceptives used at present have improved efficiency, but they have certain drawbacks that need immediate attention both from a human health and environmental perspective.Keeping in mind the importance of contraception globally, we aim to develop a safe and environment-friendly contraceptive for uterus owners by utilizing commensal bacteria of the fallopian tubes. Using synthetic biology, we plan to equip the bacteria with a progesterone sensing system that regulates the production of ovastacin, an ovum-specific human protease. Ovastacin acts as a contraceptive molecule by specifically targeting the outer layer of the ovum to prevent sperm entry. The bacteria is also engineered with a xylose inducible kill switch to ensure reversibility of the contraception and a blue-light inducible kill switch to kill the bacteria upon any environmental release.OviCloak aims to offer its users control over their fertility and uphold their fundamental rights without compromising their health or the environment.
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USP-Brazil
Title
Let.it.bee.

Abstract
Bees are very important for the environment and economy since they are responsible for the pollination of about 70% of crops worldwide. Some crops are completely dependent on bee pollination to exist. Since the 1990s, registers show a decrease in bee's populations, associated with multifactorial causes, such as the Colony Collapse Disorder and Acute Intoxication. One of the most worrying causes is the misuse of neonicotinoids pesticides, of which the most used is Imidacloprid. Bees when in contact with contaminated pollen take it to the colony, spreading the toxicity and causing mass deaths. To address this we developed a plant (Solanum lycopersicum) that can metabolize Imidacloprid specifically in pollen. We associated a pollen specific promoter, pLAT52, to the expression of the enzyme CYP6G1, responsible for metabolizing Imidacloprid in the pesticide resistant Drosophila melanogaster. This way pollen remains safe for pollinators while the rest of the plant is protected against pests.
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Linkoping Sweden
Title
CyaSalt - A novel synthetic biology solution to the global freshwater crisis

Abstract
The world population is consistently growing and integrated agriculture is expanding consequently. As a result, the global need for freshwater is greater than ever and it continues to increase. Accordingly, the world is facing a freshwater crisis that is vastly affecting the agricultural industry in all parts of the world. CyaSalt is an innovative approach to solve this crisis. The aim of the project is to desalinate seawater in an environmentally friendly way using modified phototrophic organisms. These organisms will utilize sunlight to activate the inward-directed chloride pump, Halorhodopsin, that imports chloride ions. Sodium ions will enter via the ion channel MscL. Thereafter, the modified organisms are separated from the desalinated water by a cellulose filter. The organisms bind to the filter via a carbohydrate-binding domain on their surfaces, resulting in desalinated water free from modified bacteria. Hence, CyaSalt provides a sustainable and economically accessible freshwater source for agricultural use.
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Ecuador
Title
AgroBactory 593: a bacterial platform for producing specific biopesticides

Abstract
Plant diseases are one of the main hazards to agriculture. One out of three Cavendish bananas being eaten every day is produced in Ecuador. Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (FOC) is the major threat to banana cultivars worldwide. AgroBactory 593, is a bacterial platform that produces low-cost biopesticides to fight against plant diseases. By producing and delivering double-stranded RNA molecules as biopesticides, AgroBactory 593 uses RNAi technology to silence expression of specific targets in the pathogen genome. To demonstrate the platform's effectiveness, we made a biopesticide against FOC-TR1 in vitro. Its modular design allows us to easily change the pathogens of interest, adjust the biopesticide level, and select the bacterial chassis. This way, we can optimize and customize our biopesticide providing an accessible solution for farmers and their crops against pests. AgroBactory 593 is a powerful tool to guarantee disease-free and high-quality food for the world.
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NEFU China
Title
G-quadruplex-directed colorimetric virus detection system

Abstract
The coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) severely struck the whole world. Rapid and convenient detection of pathogenic viruses can facilitate early detections and prevent large outbreaks of viral epidemics, including COVID-19. Here, we developed a G-quadruplex-based, highly specific, expedite and economic RNA virus detection system using SARS-CoV-2 as an example. The procedure includes recombinase-polymerase-mediated viral sequence amplification, nCas9-Phi29-mediated DNA nicking and displacement, rolling cycle amplification of G-quadruplex, and color development. All reactions can be carried out at ambient temperatures with limited requirement of instrument, which allows the method to be used under all circumstances.
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SZ SHD
Title
Alpha Gel

Abstract
Our goal is to be able to produce a product that can safely degrade the keratin in hair while easily removing body hair, hence solving the problem of keratin waste disposal. We have found keratinases that can decompose the α-keratin in human hair. By incorporating it into a bio-gel, we can produce a product the market has never seen before. We hope that our products can help everyone, and we hope that in the future, everyone can choose our products freely. Of course, we accept all kinds of beauty, hairless or not. As long as we like it, we should advocate social acceptance and respect the aesthetics of all people. In the future, we hope that this technology can be further applied in medical and industrial fields, such as treatment of skin keratinization diseases, treatment of poultry feathers in farms, leather pretreatment, hair treatment of household sewage pipes, etc.
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NEU CHINA
Title
Environmental multivirus detection based on PmrCAB and LuxI/LuxR quorum sensing system.

Abstract
In this study, we developed an environmental multivirus detection device based on engineered E. coli to capture coronavirus, such as SARS-CoV-2, MERS-CoV and HCoV-229E, to spark fluorescence reporter gene. The PmrCAB system derived from Salmonella was recombined onto the surface of E. coli, and the Fe(III) recognition site of PmrB was replaced with the receptor of targeted coronavirus. Therefore, the bacteria can capture coronavirus via binding to their receptors and transport signaling to the downstream components. LuxI/LuxR quorum sensing system from E. coli and Hrp amplifier from Pseudomonas syringae were used to enhance the sensitivity and output signal. In addition, we also develop an improved air sampler to enrich nanoparticle in air onto engineered E. coli. The implementation of the environmental multivirus detection would be helpful for establish a preventive system in public place, such as hospital and super market.
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GreatBay SCIE
Title
ONCOKILLER

Abstract
Breast cancer is one of the most serious health issues faced by human society. However, its solution remains flawed in many ways, for example, antibody-drug conjugates often cause great side effects; pathway inhibitor drugs rely on specific pathways which sometimes can be bypassed by tumor cells. To tackle these issues, we developed a new drug delivery system consisting of aptamers, nanoparticle, and a model drug—similar to that of ADC, but applying aptamers rather than an antibody. Moreover, we make modifications throughout the system to further improve the drug's specificity and stability: our aptamers are pH-sensitive, only binding to cancer cells in an acidic tumor environment together with the nanoparticles. We have communicated with experts and modified our project design based on their advice. In addition, we produced card games and documentaries and delivered lectures to increase our project's publicity and promote the significance of synthetic biology in the real world.
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DUT China
Title
Artificially designed delicate modular enzymes system for enhanced PET plastic degradation at mild temperature

Abstract
Wide spreading and utilization of plastic polyethylene terephthalate (PET) in the world has caused serious environmental challenges and gained much attention. In response, microbe Ideonella sakaiensis was reported to be capable of secreting two efficient enzymes to deconstruct PET polymers in mild temperature. However, this two-enzyme system degradation capacity is highly limited by inhibition effects, diffusion of intermediates and PET surface physicochemical properties. Therefore, we design a delicate multicomplex enzyme system, in which short peptide tags (RIAD and RIDD) are applied to create scaffold-free modular enzymes assemblies. In order to effectively degrade microplastic PET particles, we construct enzymes of IsPETase and MHETase and protein hydrophobin in our complex system via scaffold modular part, which reveal higher catalytic efficiency in mild temperature. This work presents an innovative strategy to improve PET degradation via biosynthetic factories and artificially designed proteins system that do not exist in the nature.
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Thrace
Title
Salica: A device for early screening of colorectal cancer using salivary biomarkers

Abstract
Colorectal cancer has high incidence across the world and the 5-year survival rate for people with a localized stage colorectal cancer is 90%, but only one third of patients are diagnosed at such stage. This problem persists despite the existence of many screening solutions such as colonoscopy and stool based tests. Our proposed solution is Salica, a non invasive, easy to use home device, that accurately quantifies salivary colorectal cancer biomarkers such as specific strains of Fusobacterium nucleatum and microRNAs. Firstly, the biomarkers are isothermaly amplified using recombinase polymerase amplification and the produced nucleic acids are targeted by the cas12a enzyme. Upon target recognition a fluorescently labeled ssDNA probe is cleaved by cas12a resulting in a colorimetric output that is quantified by a 3d printed smartphone fluorimeter. In the future our system can be adapted to any improved biomarker just by changing only the RPA primers and the cas12a crRNA.
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Bielefeld-CeBiTec
Title
P.L.A.N.T. Plant-based Ligand Activated Noxious agent Tracker - make the invisible visible

Abstract
As an invisible threat to the environment and human life, remnants of chemical weapons from both world wars still contaminate the soil. In Germany alone, there are over 200 suspected locations. We develop a plant-based detection system for degradation products of chemical weapons that is highly specific and allows cost-efficient screening of large areas while being easy to use.Our plant indicates the presence of toxic chemicals by changing its color to red. For this, we introduce two new reporter systems called RUBY and ANTHOS, enabling the synthesis of the plant pigments betalains or anthocyanins, respectively. If the chemical is present, it is specifically bound by a receptor, which then activates a signaling cascade, resulting in the synthesis of the pigments. Both computational design and site-directed mutagenesis are used to design new receptor proteins.In the future, our plant allows the detection of further chemicals by replacing the specific receptor.
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GO Paris-Saclay
Title
EndoSeek, detecting endometriosis

Abstract
Endometriosis is a painful and poorly known pathology caused by proliferating uterine-lining cells outside of the uterus. It affects approximately 10% of women worldwide and can take up to 8 years to be diagnosed. Some blood-circulating microRNAs could constitute promising biomarkers as their concentration differs between patients with endometriosis and healthy women. iGEM GO-Paris-Saclay project “EndoSeek” aims to develop a new diagnostic tool to detect these miRNAs, exploiting the nucleases Cas13a and Cas14a1 and the production of a fluorescent signal that could be measured with a smartphone. We modeled the signal amplification provided by our nuclease cascade and attempted to identify new miRNAs endometriosis biomarkers from a public database using artificial intelligence. We created a video game to inform adults and children above 10 about endometriosis and the possibilities opened by synthetic biology. Finally, following our dialogue with patients and doctors, we questioned the ethical implications linked to diagnoses.
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St Andrews
Title
The Overproduction of Shinorine (a UV-Protecting Molecule) and its Potential Application to Probiotic Sunscreens

Abstract
Chemical toxins such as octinoxate and oxybenzone, found in many commercially-available sunscreens, contribute to coral reef degradation. To combat this problem, a solution was designed whereby Shinorine, a natural, UV-protecting compound could be synthetically overproduced by probiotic Nissle 1917 E. coli - these bacteria would be combined with a lotion containing a carbon-rich nutrient medium, creating a sustainable, ocean-friendly sunscreen alternative that could be applied directly onto the skin. The aim during the Phase II stage was to clone constitutive genes into plasmids, transform these into BL21 E. coli cells, and finally test for the expression of the enzymes of the Shinorine-producing pathway. ATPG, the initial gene in the Shinorine-producing pathway, was successfully cloned into the pETDuet-1 plasmid, however cloning the other three genes into their plasmids proved difficult. To conclude, it was shown that ATPG can be successfully cloned into the pETDuet-1 plasmid via Gibson assembly.
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