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iGEM2021全チームプロジェクト概要 ~第四弾~

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四弾になります。(全四回の第四回)。
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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iGEM2021全チームプロジェクト概要 ~第四弾~
[翻訳版] iGEM2021全チームプロジェクト概要 ~第四弾~


※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。

データの見方

(例) チーム名(チームページリンク付き)
タイトル
要約
Wikiへのリンク

Shanghai Metropolis
Title
EV71 oral vaccine against HFMD

Abstract
Hand-foot-and-mouth disease (HFMD) is a contagious viral infection, mostly occurring in children under the age of five. The condition is caused by a series of enteroviruses, including EV71, and VP1 is one of its pathogenic subunits. Since many of the infected children may be afraid of traditional injection-based vaccines, we are trying to develop an EV71 oral vaccine that will gain higher compliance among infant patients. Using E. coli and L. casei as the expressive system, we express the plasmids we constructed, pGEX-6P-1-VP1 and pGEX-6P-1-VP1-LTB. We hope that our project will shed light on the possibility and practicability of the EV71 subunit oral vaccine and pave the way for a much safer, more convenient, and efficient oral vaccine for children to defend against HFMD.
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Shanghai Metro Utd
Title
In vitro production by E.coli of Angiogenin and the potential in curing neurodegenerative diseases

Abstract
Parkinson's disease (PD) is an inflammatory brain disease that affects 20 million people worldwide and the incidence is expected to rise. However, current medical treatment is not optimal, and thus an effective treatment is very much needed. Surprisingly, we discovered that a specific protein called Angiogenin is reported as a potential therapeutic target to neurodegenerative disease given. In this project, our team Healers on Vessels aim to construct a recombinant E. coli by biomedical engineering that can continuously synthesize functional ANG protein, which could possibly serve as an in vitro method to massive produce ANG for clinical use.
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ASU
Title
Genetically engineering the microalgae Chlamydomonas reinhardtii to sequester arsenic from contaminated groundwater

Abstract
Arsenic contamination in groundwater is a serious problem both in local Arizonan communities and abroad: prolonged exposure to arsenic contamination can cause cancer, vascular damage, and liver failure. This project aims to engineer the microalgae Chlamydomonas reinhardtii to sequester arsenic out of water. Metallothionein, arsenate reductase, and ferritin were integrated into the microalgae via the pASapI plasmid in varying permutations. The plasmid rescues function of the photosystem II gene, leveraging the ability to photosynthesize as a selective trait. Metallothionein and ferritin bind the two most common forms of arsenic: arsenite and arsenate, respectively. Arsenate reductase catalyzes the reduction of arsenate to arsenite, allowing for the ultimate sequestration of the toxic metal to occur in the chloroplast. Transformed algae were incubated with multiple concentrations of arsenic-contaminated media and the final concentration of arsenic after 2-3 days of exposure was measured using ICP-MS to quantify uptake efficacy.
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Bolivia
Title
ARSEMAPHORE

Abstract
Reliable access to potable water is a major problem in Bolivia and other nations. Scarcity of this essential resource drives populations to seek drinking water from rivers, lakes, and wells. However, these sources can be dangerous if they are contaminated by heavy metals. Preliminary studies show that drinking water in Bolivian cities often contains highly hazardous levels of arsenic. In response, we designed and constructed a bacterial biosensor to rapidly quantify water sample arsenic content. The biosensor design strategy is called traffic light; it employs 4 genetic constructs to quantify defined ppb ranges of arsenic content in water. Light color responses indicate lower metal concentrations while increasingly dark color responses correspond to increased arsenic content. We created a portable device that houses the biosensor and allows one to quantify water samples anywhere. Our goal is to make the biosensor available to users in Bolivia and eventually around the world.
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Purdue
Title
Argonaute Mediated Viral Point of Care Diagnostic Device

Abstract
The COVID-19 pandemic has strained global diagnostic capacities and highlighted the limitations of conventional lab-based assays. Current on-site kits have false-negative rates as high as 33%. In an effort to provide accurate, non-invasive, affordable, and rapid Point of Care(POC) diagnostic tests for COVID-19 and other emerging pandemics, Purdue iGEM since the summer of 2020 has been working on cArgo: an Argonaute mediated microfluidic viral diagnostic device. cArgo extracts viral RNA from saliva for amplification and conversion into dsDNA. The TtAgo Argonaute protein then cleaves the dsDNA using viral strain-specific DNA guides producing ssDNA fragments. These fragments serve as secondary guides allowing the Argonaute to cleave the molecular beacon emitting a quantifiable fluorescent signal for conclusive result determination. Coupling the biologics with chip barcoding and app integration, we hope to revolutionize POC Diagnostics while making data more accessible for simultaneous detection and contact tracing.
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Stanford
Title
Mitigation of Grapefruit-Drug Interaction through Engineered Gut-Colonizing Yeast

Abstract
The grapefruit-drug interaction refers to unpredictable adverse effects that occur when certain drugs are taken concurrently with grapefruit products. Grapefruits contain a family of compounds called furanocoumarins, which are known to repress CYP3A4, an enzyme involved in the metabolism of a large percentage of drugs prescribed today. This interaction causes increased bioavailability of some drugs and can result in unintentional drug overdose. Given the prevalence and popularity of grapefruit products and the number of affected drugs, our project is designed to provide passive negation of the grapefruit-drug interaction. Our plan is to put the gene for CYP6B1, an enzyme from the black swallowtail butterfly that breaks down furanocoumarins, into brewer's yeast and colonize the human gut microbiome with our modified microbe to provide continuous mitigation and resilience to the drug interaction through the gene's expression.
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McMaster
Title
Genetically Modified Bacteria for Targeted Elimination of Adherent-invasive Escherichia coli in Inflammatory Bowel Disease

Abstract
Inflammatory bowel disease (IBD) is a group of immune-related diseases, characterized by chronic inflammation and severe tissue damages in the human gastrointestinal tract. Therapeutics developed using synthetic biology methods showed some efficacy in laboratory studies but failed to produce clinically relevant results. In this project, we performed a comprehensive literature review of recent advancements in the field and we proposed the use of bacteriocin to target the pathogenic bacteria adherent-invasive Escherichia coli (AIEC), which are closely involved in the development of IBD. We constructed a genetic circuit that can simultaneously detect the inflammation marker nitric oxide, and a pathogen-specific quorum sensing factor named autoinducer-3. Upon activation, the genetic circuit expresses colicin E1 and E9, two bacteriocins that can selectively eliminate the AIEC. We designed detailed experimental workflows for circuit construction through Gibson assembly and for functional verification. A mathematical model was also established to predict the outcomes of our design.
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LINKS China
Title
NeoLeathic Age — Revolutionary Leather Substitute from Kombucha, Spider-silk, and Natural Dyes

Abstract
The booming of the fashion industry has led to rising demand for leather. However, current industrial leather production causes environmental and ethical problems such as water pollution and animal cruelty. LINKS_China 2021 aims to create a sustainable and humane leather substitute using bacterial cellulose (BC) produced by co-culturing Komagataeibacter with Saccharomyces cerevisiae. By engineering S. cerevisiae, we induced transgenic expression of artificial spider-silk proteins fused with cellulose-binding matrixes, which increases the tensile strength and flexibility of BC membrane upon binding. Additionally, we enabled our engineered S. cerevisiae to synthesize ethyl acetate, giving our membrane a fruity fragrance. To endow our leather with more fashionable features, we used engineered Escherichia coli to synthesize different di-halogenated indigoid dyes for coloration. We expect our novel leather to be a transformative product that will continue the prosperity of fashion industry through a more humane and environmentally friendly technology.
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AISSU Union
Title
Memory bread:Constructing Catechin Production Pathway in Saccharomyces Cerevisiae

Abstract
Increasing ageing of population has become a global issue, and the cognitive ability of those seniors, an important measurement of this group's healthy level. Researches have proven that flavonoids could be potential in improving the seniors' cognitive ability. We selected nine different genes from plants to build up two plasmids and used Saccharomyces cerevisiae as the chassis for expression. The first plasmid uses L-Tyrosine to produce naringenin, a central molecule in the production of flavonoids. The second plasmid converts naringenin into the final product -- catechin. As a crucial member of flavonoids, catechins plays a significant role in improving cognition and cardiovascular health. We hope that catechins could be used as food additive in elders' daily diet, and therefore improve their memory. And we believe that it has the potential to become a new solution towards seniors' health in the future.
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Whittle
Title
Mural artist: Cyanobacteria-produced Auxin to Engineer Plant Growth in Horticulture

Abstract
Plant hormone plays an important role in agriculture and horticulture manufacture. As such, they have gained widespread commercial use in nurseries and in farming. This year, Whittle iGEM tries to produce indole-3-acetic acid (IAA), a main plant hormone, by using Cyanobacteria PCC6803. First, we introduced the gene of yeast into Escherichia coli, and construct the IPA pathway. By taking Trp as the substrate, we verified that the genetic pathway is able to convert 80% of Trp into IAA within 48 hours. Afterwards, we built the expression vector of cyanobacteria and convert it into Cyanobacteria PCC6803. We hope to manufacture cyan sponge bricks that contain this microorganism to produce our product Auxwall, and promote the growth of plants in garden landscaping. We hope that this product will transform the agriculture and landscape design in the future.
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SCAU-China
Title
MESEG

Abstract
Heavy metal pollution is very serious in water. Our team came up with the idea of a "cruisable molecular magnet". "Molecular magnet" means it can capture heavy metal ions and store them in vacuoles. This can be achieved by expressing heavy metal binding proteins and autophagy-associated proteins in Chlamydomonas reinhardtii. And "cruisable" means it can swim back and forth in sewage by our hardware device. This new device have an acrylic frame and cruise system, supplemented by sodium alginate embedding method and semipermeable membrane. On the one hand, it can cruise in the water by Beidou navigation satellite system (BDS) or remote control to capture heavy metal ions at different positions and facilitate recovery at the same time. On the other hand, it can prevent the leakage of genetically modified Chlamydomonas and provide a relatively stable and safe environment for Chlamydomonas.
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KEYSTONE
Title
Rubber Eater———on the biodegradation of rubber (cis-1,4-polyisoprene)

Abstract
In the enormous yet growing global rubber production, we see an urgent issue: waste rubber. By interviewing rubber recycling managers in China, we found that only 7% of 12945000 tons of rubber is recycled in 2020, meaning most rubber waste has either entered a landfill/open dump or an incinerator, both entail serious ecological/safety hazards. To combat this issue, we employed Latex Clearing Protein (LCP) as a means of rubber biodegradation. However, the implementation of LCP still faces many obstacles. To facilitate real-time rubber degradation, we introduced a signal peptide named hlyA that allows for the extracellular secretion of LCP, and for water solubility, we added a fusion protein named NusA. In addition, the signal peptide provides our project the potential for sustainable industrial-scale use. With improved efficiency, this project will become a promising and environmentally friendly rubber waste management method, hopefully contributing to the solution of this pressing global issue.
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NUDT CHINA
Title
Light-mediated control of cell cycle in mammalian cells

Abstract
Optogenetic tools provide essential approaches to control designer cell function. Our project intends to optimize Predator Pro, a modularized protein degradation system we've been working on in the past few years, to achieve light-mediated degradation of endogenous cyclin protein, the key regulator protein family in cell cycle, thereby manipulating cell cycle in mammalian cells. Integrating with a raspberry pie-based blue light illumination device, our system may provide novel tools for cell cycle synchronization and spatiotemporal control of cell division. Our work may provide novel tools for synthetic biology community and researchers in other fields, it will also provide new hints for treating diseases caused by cell abnormal division.
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NOVA LxPortugal
Title
P(L)AST: The Ocean Cleaner of the Future

Abstract
Microplastics are a growing concern worldwide. Studies point to about 5.25 trillion pieces of plastic floating in our oceans, and our society is only now beginning to grasp the extent of the health risks associated with microplastic consumption. All these concerns led us to develop P(L)AST, a simple solution to battle this ongoing crisis. By transforming Bacillus subtilis with the Polyethylene terephthalate (PET) degradation pathway present in Ideonella sakaiensis, we aimed to create an efficient, versatile, and easy to deploy plastic degradation platform – qualities further improved through our metabolic optimization models. P(L)AST is also a project concerned with social impact; as such, we spent a portion of our resources inspiring the younger generations and educating the masses by organizing, promoting, and participating in events related with the environment, sustainability, and science.
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TokyoTech
Title
Lacteeth

Abstract
There are billions of people who have dental caries around the world. Dental caries are related not only to quality of life but also to health expectancy. The more aging society grows, the more dental health problems become important.Our project aims to develop new probiotics that continuously prevent dental caries with engineered Lactococcus lactis. We designed Lactococcus lactis which is able to bind specifically to biofilm and diffuse growth inhibitors against bacteria causing dental caries. We also considered the practical use of recombinant probiotics in the real world and suggest how we can use engineered bacteria safely outside of the laboratory. We hope that our project contributes to the world without dental caries.
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Estonia TUIT
Title
SALSASMILE

Abstract
Around 2.3 billion people suffer from caries. The preventive measures include dietary restrictions, exposure to adequate doses of fluoride, and tooth brushing. However, inappropriate tooth brushing may cause dental cavities and harm gums. Therefore, safer and unified oral hygiene methods are in demand.Tooth decay is caused by bacteria that bind the human SALSA protein on teeth' surface, forming a biofilm and producing acids that destroy teeth. In SALSASMILE, we engineer a protease to target the SALSA protein. We expect that cleaving SALSA will remove bacteria even from the inaccessible areas, making oral hygiene more efficient. We develop a simple assay in yeast to select for proteases that cleave the desired sequence. Combined with deep mutational scanning or directed evolution, this is a powerful tool to engineer proteases.Everyone, especially children, people with orthodontic cases, or disabled individuals could use SALSASMILE alongside tooth brushing for improved oral hygiene.
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Washington
Title
RhizoMet

Abstract
Although centuries of technological advances in manufacturing and industry have improved humanity's standard of living, they have often come at the expense of environmental well-being. The industry of mining, smelting, and refining heavy metals often leaves behind hazardous waste in the soil and air. Our project, RhizoMet, addresses these toxic levels of arsenic in soil through the use of bioremediation. As our proposed proof of concept, we plan to test the binding efficacy and specificity of our modified metallothionein proteins to arsenic. To accomplish this, metal tolerance and accumulation assays were performed on E.coli overexpressing these proteins. Given an effective model, we plan to incorporate these E.coli into the roots of Lomatium triternatum and other RhizoMet plants, remediating arsenic-polluted areas like those around metal smelteries. Through future iterations of our RhizoMet plant-E.coli system, we hope to expand the impact of this bioremediation method for areas like Northport and beyond.
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CHS Missouri US
Title
AquaeSCOPE: Investigating our Water Distribution Systems

Abstract
While communities lacking access to safe and regulated water distribution need alternative water sources, a general rise in distrust and misinformation towards the microorganisms in municipal water has led many individuals to avoid safe tap water for the environmentally costly alternative of bottled water. In order to investigate this problem in our community, CHS_Missouri_US created a synthetic biology system that can examine tap and bottled water microbiomes. Utilizing shotgun metagenomic sequencing, we have built sequencing libraries targeting the V4 region of 16S rRNA genes extracted from the water samples. Unique molecular barcodes were attached to the samples to comprehensively identify the taxonomic profile and relative abundance of all genera within both samples. Research was then conducted to determine the pathogenic risk of the microbiota detected. The documentation for this process can be applied to characterize the microbiome of water sources globally and provide insight towards improved water contaminant testing.
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Lubbock TTU
Title
ChiSPY: a Chitin-Sensing Pathway for the novel detection of chytridiomycosis in amphibians

Abstract
The global chytridiomycosis pandemic caused by fungi, Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salmandoris (Bsal), is responsible for the decline and extinction of many amphibian species. Standard in-field detection methods such as MicPCR and MinION sequence internal transcribed spacer regions from swabs of epithelial cells to indicate the presence of fungal zoospores. However, these methods are expensive, time-consuming, and result in varying infection intensity across Bd strains. To address these concerns, we developed a microbial biosensor by engineering E. coli to express the chitin-activated V. cholerae one-component system coupled with a GFP reporter. Diverging from traditional detection methods that indicate zoospore presence, chitin in the cell walls of fungal zoosporangia will serve as the biomarker to indicate infection. This novel biosensor will provide field ecologists with an accessible alternative method to conduct preliminary site testing.
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Shanghai Metro
Title
Gene technology for protecting patented bacterial strains

Abstract
Protecting patent biotechnological strains is usually a costly and time-consuming process. Aiming to provide a fast, practical, and inexpensive way to protect patent rights over registered strains, our team constructed a genetic tool that disables the strains if they're replicated without authorization. This mechanism is developed based on the Type IV Secretion System (T6SS) of Gram-negative bacteria which would inject effectors to inhibit the growth of recipient cells. However, these effectors are inactive in the donor due to the presence of immune protein. In our project, the plasmid that constitutively expresses Tke (effctor) was transformed into E. coli, and the immune factor Ike is induced by tetracycline. After several tests, a suitable concentration of inducer that effectively disables the defense system was determined, which allowing the normal multiplication of the E. coli.
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CSMU Taiwan
Title
GotCha

Abstract
Upon identifying shortcomings in current detections of UTUC , the iGEM CSMU_Taiwan team aims to overcome them by developing GotCha: a precise, in vitro, and safe diagnostic kit, which also carries a potential to be employed in the detection of various micro-RNA identified diseases.
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Shanghai United HS
Title
A new engineering strain which could kill oral bacteria and prevent dental caries

Abstract
Dental caries, also known as tooth decay, have been one of the most usual diseases among people. It can directly disrupt oral health and even trigger the infection of other parts of the body. The main factor of Dental caries is the erosive organic acid created by bacteria on the surface of teeth' metabolism. Chimeric Lysin ClyR is an enzyme originally produced by Bacteriophages that specifically attack bacteria that causes dental caries. Dr. Phage's mission is to create a kind of gene engineering E.coli strain that can enter the mouth and emit ClyR when induced of arabinoxylan. With this strain, people could avoid the further erosion from those oral harmful bacteria and achieve short-term protection, long-term prevention effect.
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Gifu
Title
Trans-cleavage endonuclease specific fatigue quantification system (TESTIFY )

Abstract
Japan has often been reported as a stress-filled society in recent years. However, many Japanese neglect the excessive stress that can cause depression and other problems. We assume this is due to the difficulty of quantitative measurement of stress and that patients cannot be diagnosed by appearance whether they are suffering or not. Human Herpesvirus-6 (HHV-6) is a virus that infects 90% of humans in childhood and has the characteristic that it can remain latently in the human body for a lifetime. It has been reported that fatigue over a period of about a week reactivates the virus and is detected in saliva. Accordingly, we decided to create a system to quantify HHV-6 by using CRISPR based DETECTR assay (2018 Doudna et al.), based on our idea that the amount of the virus in saliva can be correlated with the stage of medium-term stress.
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Gunma
Title
Breakdown Biofilms in Baths

Abstract
Hot spring facilities all over the word have various problems caused by biofilms. For example, biofilms protect infectious microorganisms from disinfectants. Biofilms are communities of microorganisms that irreversibly stick to non-biological surfaces and grow on the surfaces, producing exopolysaccharides that facilitate attachment and matrix formation. To break down such biofilms, we constructed a vector for expression of a glycolytic enzyme fused to RFP (red) that degrades exopolysaccharides. An Escherichia coli strain was used not only as an expression host but also as a carrier of the enzyme to biofilms. The strain also contained a suicide vector as the second vector that can cause bacteriolysis under some specific condition. Our findings showed that experimental biofilms were efficiently degraded by the glycolytic enzyme that leaked from the lysed transformant cells . The use of the red transformant may make it easier to kill infectious microorganisms with disinfectants or remove biofilms from bathtubs.
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BHSF
Title
Synthesis of Carminic Acid Using Gene-edited E.coli

Abstract
The general idea of this experiment is to use the gene edited E.coli to express the gene fragments of polyketide synthase, cyclase, monooxygenase and c-glucosyltransferase. After enduring a series of chemical reaction, the edited E.coli is able to generate carminic acid.
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Athens
Title
AdAPTED: Augmenting dNTPs And Polymerase Through Enzymatic Design

Abstract
Nucleic acid amplification (NAA) techniques, such as Polymerase Chain Reaction (PCR), are a standard biological laboratory technique, also used for diagnostics. The aim of this project is to make NAA techniques more accessible. Two of the key components of PCR are a polymerase and the four deoxynucleotide triphosphates (dNTPs). However, the total cost of dNTPs' production is significant and they are often produced by complicated, unsustainable chemical processes, mainly feasible in centralized laboratories with expensive equipment. For this project, a high yield genetic circuit is designed in order to produce ribonucleotide reductase (RNR) and thymidylate synthase (TSase), two enzymes necessary for the production of dNTPs. The produced dNTPs, along with thermostable Pfu DNA Polymerase, also included in the designed genetic circuit, allow for a low-cost PCR, with components not requiring cold storage. This procedure allows for an alternative approach for the production of basic PCR reagents.
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EPFL
Title
CuRe: Copper Bioremediation to Revitalize Vineyards

Abstract
Mildew is a destructive fungal disease of plants including wine-producing grapevines. For more than a century, mildew has been treated with ‘bio' farming compatible copper-based fungicides which adhere to plant leaves. However, though only trace amounts of copper is present in wine made from treated plants, when rain washes the fungicide away, toxic levels of copper contaminate the surrounding soil, inhibiting new cultivation and biodiversity. Our goal is to remove copper from grapevine rainwater runoff to prevent this pollution. We generated yeast strains which express the normally intracellular copper-binding protein CUP1 in multiple copies and configurations on their surface to enable bioadsorption of environmental copper. As a proof-of-concept for a real-world implementation of our decontamination strategy, we have designed and built a prototype bioreactor to enable the removal of copper from rainwater by our modified yeast. We envision our solution could help protect wine growing regions from copper soil pollution.
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UPenn
Title
Engineering the OptoReader: a device for high-throughout optical stimulation, measurement, and feedback in microwell plates

Abstract
Optogenetic tools allow for unprecedented dynamic control, immediacy, and reversibility of metabolic processes. However, there lacks methods to stimulate and monitor optogenetically-responsive samples simultaneously in a high-throughput manner. We developed the OptoReader to address this gap. The OptoReader is an affordable, open-source device that couples optogenetic stimulation with real-time fluorescence and optical density measurements, allowing for 96 programmable parallel experiments. It consists of two circuit boards. The stimulation assembly is a modified optoPlate-96. The reader assembly is an array of photodiodes and UV LEDs compatible with a standard 96-well plate. Arduino microcontrollers control both circuit boards to communicate with a general computer and each other. We demonstrate OptoReader measurement of OD and fluorescence in bacterial cultures and show how OD readings can be used for feedback controlled optogenetic stimulation parameters. We expect that the OptoReader will enable entirely new optogenetic experiments with impact in both basic and applied sciences.
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Michigan
Title
Investigating the Versatility of Encapsulin Protein Nanocompartments for Drug Delivery

Abstract
Encapsulins (Encs) are a class of proteins that self-assemble and envelope other functional “cargo” proteins when co-expressed. Encs are relatively simple protein shells, yet they are stable and can carry robust cargo. We believe this represents an opportunity for novel drug delivery. In this work, we investigate the intake of assembled Encs by living eukaryotic cells and whether the Enc containers can be disassembled by protein-degradation enzymes native to the targeted organism. Brewer's yeast (S. cerevisiae) was selected as the proof-of-concept organism for this project, and mNeon green was chosen as the model Enc cargo. We are cloning bacteria that will produce Enc shells with yeast mating pheromone tags displayed on their surfaces. The tags will encourage the yeast to endocytose the disguised Encs into the cell from the environment. Success will be measured by using fluorescence imaging to confirm the presence of assembled mNeon green-bearing shells within live cells.
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NYU New York
Title
Flutec - Influenza Diagnostic Test

Abstract
The NYU iGEM team is currently developing an affordable, portable, paper-based diagnostic device for the influenza A strain using novel techniques such as toehold switches that can be performed at home without the need for expensive laboratory equipment. Research done over the past year revealed the impact of COVID-19 on healthcare staff and services, as well as the need for expanded home testing options for common illnesses. As a response to this need, our device creates a simple method to detect influenza A with a saliva sample that can be easily distributed to aid local and global public health efforts by increasing access to care.
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NMU China
Title
Toggle Macrophages

Abstract
Elimination of the SARS-COV-2 and balance the immune response are both important for late-stage COVID-19, however, no ideal therapeutics can achieve the two goals for our knowledge. To address this, we have 1) designed synthetic antiviral macrophages with immune-regulatory capacity and 2) demonstrated its feasibility through both in vitro experiments and mathematical modeling. The designed synthetic antiviral macrophages, termed "Toggle Macrophages", have the phagocytic capacity specific for SARS-COV-2 and showed pro-inflammatory/anti-inflammatory phenotypic transformation according to the level of local IL-6 concentration. Therefore, as "smart" therapeutics, Toggle Macrophages can both reduce the viral load through phagocytosis and balance the immune system via phenotypic transformation. We further mathematically modeled and analyzed the feasibility of the practical application of Toggle Macrophages in humans. This work supports the use of Toggle Macrophages as a novel Genetically Engineered Cellular Platform that may be useful in future therapeutic approaches against severe COVID-19.
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HKIS
Title
Vib-Check

Abstract
Vib-Check is a two-part detection and purification system for Vibrio bacterium in Oysters. Vibrio, being the leading cause of seafood-related deaths around the world, is a hazard that has yet to be tackled effectively. Vib-Check aims to provide a method for affordable and effective detection of Vibrio pathogens in oyster batches that the average fisher can use, something no product is currently capable of. In the event of a positive result, aquaculturists can use our specially designed peptides expressed by transgenic microalgae for the effective and convenient purification of pathogens in oysters.
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United Shanghai HS
Title
PET Gig Eater

Abstract
Plastic waste pollution has seriously threatened the environment and creatures. Here, we designed a burying yard with concentrated PET plastics, using the modified Bacillus subtilis carried by earthworms to initially degrade PET ,accelerating the degradation of PET plastics in it. At the same time, the activities of earthworms are used to improve the distribution of engineering bacteria and enzymes,solving the difficulty of frequent mixing in the dump site.The modified engineering bacteria release PETase and MHETase to hydrolyze PET into ethylene glycol and terephthalic acid,finally metabolizing it into carbon dioxide and water. What's more ,we also inserted GFP gene into the plasmid to further track the distribution of engineering bacteria in the soil. And by detecting the fluorescence in the body or feces of the earthworms, we can know whether the engineered bacteria have been swallowed by the earthworms. The expected result is that the engineered bacteria can be carried elsewhere through the activities of earthworms, which will promote the distribution of engineered bacteria in the soil.
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LMSU
Title
ASCEND

Abstract
The question of food supply for the long-distance flights remains still unsolved. Moreover, onboard food production must meet restricted requirements due to the curtailed resources on the spacecraft. Сyanobacteria Arthrospira platens is a perfect candidate in this case. However, it is tasteless. A long-term ASCEND project is aimed to introduce Arthrospira platensis engineered to produce any genes of interest, and flavours in particular, as a new chassis for the synthetic biology community. We have designed and tested a special optogenetic system, which will facilitate switching between different products and help optimize growth and production conditions. Blue light induces anchoring of BcLOV4 protein in the plasma membrane and maintains culture growth, whereas far-red light induces the production of genes of interests by activating the BphP1 light-sensitive protein and forcing it to inactivate QPAS1-Gal4 repressor. This year genetic constructions were trialed in E.coli with YFP as a test gene.
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NOLA
Title
Site directed mutagenesis of CRISPR/Francisella novicida Cas12a system to enable multiplexing

Abstract
Multiplexing of the CRISPR/Cas system increases gene-editing efficiency and expands CRISPR/Cas function to multiple locations at once. However, multiplexing involves using repetitive sequences in the crRNA arrays, which can be unstable on plasmids and susceptible to recombination. We are designing new sequences of the crRNA scaffolding region of the CRISPR/Francisella novicida Cas12a system to expand the crRNA sequences available for multiplexing. We performed site-directed mutagenesis by PCR on our crRNA scaffold sequences, created linear DNA amplicons comprising multiple mutations each, and ligated the ends to make a circular plasmid. We transformed the recombinant plasmid libraries, which included a nuclease dead Cas12a (dCas21a) into E. coli with functional plasmids inhibiting the promoter sequence of pGEX-iLOV (called CRISPR interference, or CRISPRi). Using fluorescence activated cell sorting (FACS), we sorted out the functional plasmids. We will analyze next generation sequencing data to identify and characterize the successful sequences.
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Duke
Title
NODES: Novel Organoid-dependent Drug Efficacy Screening implementing high-throughput, droplet-based brain organoids

Abstract
Clinical treatment of glioblastoma, a disease that kills over 10,000 Americans annually, is limited by the lack of a scalable, physiologically-relevant model for testing therapeutics. Duke iGEM is developing NODES, a high-throughput organoid-based drug screening platform to characterize treatment efficacy in common glioma variants. We designed a non-invasive reporter device that quantifies the drug response of mutation-specific glioma cells in a mini-brain co-culture model, grown in a droplet-based system. Additionally, we modeled our reporter system, which detects oncometabolite levels throughout brain tumor development, to improve device characteristics and developed a machine-learning based image analysis pipeline for organoid screening. To identify the social and ethical implications of our work, we interviewed patients, clinicians, and other stakeholders and integrated their feedback into our design. By recapitulating the brain microenvironment, NODES has the potential to accurately characterize drug responses, offering new hope to patients in their fight against this lethal disease.
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MiamiU OH
Title
CROP: Creating RuBP Optimized Photosynthesis

Abstract
Global agricultural productivity is projected to not meet the needs of increasing populations developing higher standards of living. On a cellular level, crop yield is limited by the inefficiency of photosynthesis. Our project aims to improve this efficiency by implementing an alternative RuBP regeneration portion of the Calvin-Benson-Bassham (CBB) cycle. Two alternative pathways, which use enzymes from other reactions that act on common metabolites used in the CBB cycle, were explored first via computational modeling. Impacts on growth and reaction fluxes in silico assessed the validity of these pathways in creating a more robust photosynthetic cycle. One of these pathways which overexpresses the native enzyme transaldolase, was also assessed in vivo. Ultimately, we showed the validity of two alternative pathways in allowing a more efficient regeneration stage of the CBB Cycle. These pathways could eventually be implemented into higher plants to allow more robust cycling and therefore higher crop yield.
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MEPhI
Title
MISS - Microbiomic Space Suit: Synthetic Human Microbiota to Stimulate Radioprotection to Ionizing Radiation

Abstract
Project MISS is aimed to create a Microbiomic Space Suit protecting cosmonauts, astronauts, and taikonauts from aggressive radiation during their long-term missions. Microbial communities cover the human body and form a biological suit that helps humans to perform various vital functions. The project aims to populate human's natural “suit” with gene-modified bacteria synthesizing radioprotective proteins. New microbiota will pave a way for further space exploration plans including human settlements on the Moon or an expedition to Mars.We will modify Human bacteria using silicon nanoparticles with absorbed genetic material. Inserted genes will motivate bacteria for secreting radioprotectors packed in Outer Membrane Vesicles (OMVs) capable of fusing with human cells. The bacterial communities developed within this project will be the first synthetic human microbiota engineered to perform specific functions in its natural epitopes.
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Victoria Wellington
Title
Tropane alkaloid biosynthesis in prokaryotes

Abstract
Tropane alkaloids are plant secondary metabolites and include important medicinal compounds. Most applications are related to neurochemistry and range from the treatment of neuromuscular disorders, including Parkinson's disease, to the use as stimulants. There is increasing need for large-scale, climate-independent, and local production of tropane alkaloids as precursors for medicinal drugs. Our goal is to remedy the impact of world crises on the cultivation and exportation of these drugs. To this avail, we aim to develop a biosynthetic route for a tropane alkaloid intermediate, tropine, in Escherichia coli and the cyanobacterium Synechococcus elongatus. To the best of our knowledge this would be the first production of tropine in a prokaryotic organism and could provide an effective and cheap alternative to current tropine production methods.
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MTU-CORK
Title
Bioremediation of MARS using engineered bacteria in a novel housing system, the Biofrag 2.0

Abstract
Mars holds great interest to humanity and commercial applications alike due to its suitability for potential habitation. One issue withstanding are the components of the planets soil that contains toxic levels of sodium perchlorate making growth of food impossible. Our team has devised a solution by engineering E.coli with genomic islands from A. suilum which breakdown the sodium perchlorate in the soil through the enzymes Perchlorate reductase and Chlorite dismutase.The bacteria is housed within a 3D printed, novel, spherical design allowing biofilm formation, bacterial proliferation, enzyme distribution and addition of a seed for bioremediating and cultivating the Martian soil. This design has been titled the Biofrag 2.0.The Biofrag can be used within specially contained Biofrag Isolation Units which allow for contained remediation of soil whilst preventing widespread contamination. Each box also allows for optimal conditions for growth of both flora and microbiome within the Biofrag.
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RHS-Calgary
Title
GlucoLitter: A colorimetric biosensor for detecting glucose in feline urine

Abstract
The purpose of our project is to address the issue of effectively revealing signs of feline diabetes, as current solutions are often limited and time-consuming. Our primary goal was to produce a biosensor that creates a purple pigment in the presence of elevated levels of glucose which will act as an indication of feline diabetes. A reaction will occur between the glucose in a cat's urine and oxygen, catalyzed by glucose oxidase. This will produce hydrogen peroxide, which will react with a colorimetric assay to produce a color change. Through synthetic biology principles, we aim to apply our biosensor to provide cat owners with an early detection method for feline diabetes and other conditions.
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NTU-Singapore
Title
Rapid Detection of Single Nucleotide Mutations in COVID-19 and Cancer

Abstract
As we have learnt from interviews, assays that can detect mutations accurately are valuable in the diagnosis of fast evolving infectious diseases and cancer. Furthermore, in pandemics such as COVID-19, rapid point-of-care detection is essential to isolate patients as fast as possible and minimize spreading of the disease. Hence, we have developed a sensitive and easy-to-use diagnostic kit that can accommodate direct patient samples without RNA extraction and deliver results in under 40 minutes. Loop-mediated isothermal amplification (LAMP) was utilised to amplify the target genes. After amplification, a novel CRISPR-Cas12a Lb-RVRR enzyme was employed to identify different mutations based on the fluorescence intensity. Alternatively, we developed a probe that was sensitive to point mutations, allowing us to distinguish between different genetic variants in COVID-19 and cancer. Thus, our project has provided with better methods to diagnose such diseases for a better future.
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XJTLU-CHINA
Title
Dr. Phage

Abstract
In summary, our project aims to quantitatively detect the bacterial pathogens in food by using the edited assembly-deficient phage to target and lyse the bacteria, and then release the exogenous protein LuxR; the released LuxR can then activate downstream cell-free gene circuit and generate visual signals. The CRISPR-Cas9 system is applied to disrupt gp24 gene, an essential gene for T4 phage assembly, with LuxR gene to produce LuxR as signalling molecules for downstream system activation and prevent phage assembly at the same time. The assembly-deficient phage cannot therefore replicate in the host cells more than one generation. The downstream circuit outputs binary signals indicating whether the concentration of bacteria exceeds the national standard or not: if the concentration of the target bacteria is higher than the standard, the system will output a visual signal, otherwise, the signal will not be displayed.
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Yucai SZ
Title
A composite material for fresco restoration based on engineered E.coli

Abstract
The restoration of traditional frescos is a difficult process. Our project is to operate the modified E.coli which can be induced by blue light to produce a layer combined calcium carbonate, pigment and adhesive directly at the damaged position of the fresco. After the combination of these three, a micron level colored calcium layer will be formed on the wall surface, so as to repair the damaged frescos.
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IOANNINA
Title
Antibyeotic: Bacteria against AMR

Abstract
The increasing amount of antibiotics in the environment has led to a rising number of antibiotic resistant bacteria, putting human and animal health at risk. Antibiotic use in poultry farming remains high and leads to antibiotic pollution. The problem of AMR is now a serious threat to the achievement of many of the Sustainable Development Goals as indicated by the United Nations. Defending the One Health approach, we aim to design a bacterial mechanism, which can inactivate tetracyclines and macrolides in the surrounding environment, while containing a self-destruction mechanism, activated when it has fulfilled its role. For this purpose, we partnered with iGEM IISER-Tirupati, to share their kill switch based on the action of a modified bpDNase1. The bacterial cell death is regulated by the presence of antibiotics using TetR and MphR as antibiotic sensors. Our idea could be applied in chicken manure making the final product safe and antibiotic-free.
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Guelph
Title
CRISPR to Combat Climate Change

Abstract
Our project this year aims to address the threats posed to global food production by climate change in the form of abnormal or unseasonal weather events. Using a CRISPR activation (CRISPRa) system in conjunction with a molecular switch, we aim to create genetically modified plants for which specific stress-response genes can be manually upregulated following the human-mediated input of a chemical stimulus.Our team has also developed and executed a strong outreach plan. We have worked with professionals in the industry and experts in the fields of agriculture and genetic engineering in order to better understand how our research can be applied. We have also partnered with other iGEM teams to create a series of academic materials covering different facets of synthetic biology. We have partnered with teachers in order to gain feedback on this work, and to determine how this material could be effectively deployed in a classroom setting.
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SDU-Denmark
Title
PsiloAid

Abstract
We want to contribute to the utilization of psilocybin's absolute potential by reducing the costs of acquiring the substance for research projects. Psilocybin is currently undergoing clinical trials, to explore its potential as a treatment method for depression. However, it has shown itself to be a very difficult and expensive process to chemically synthesize psilocybin or purify it from the mushrooms. Despite recent improvements in heterologous production of psilocybin using either bacteria or yeast, current methods persist to be tedious and costly with a reduced yield. That is why we aim to construct an enhanced biosynthesis of psilocybin from tryptophan, by introducing all four different enzymes from the Funghi in Escherichia coli. After successfully producing psilocybin in E. coli, we hope to transfer this system to our ideal, non-pathogenic host organism, Rhodovulum sulfidophilum that only needs sunlight and seawater to grow, thereby making a sustainable and safe cell factory.
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NYCU-Taipei
Title
Natto It Out

Abstract
A fatal cardiovascular disease, Deep Vein Thrombosis (DVT) is a rising major health problem due to the potentially lethal complications such as pulmonary embolism. Since an ounce of prevention is worth a pound of cure, our project “Natto It Out” aims to prevent DVT progression at its earliest onset via a fibrinolytic protease—Nattokinase. In this project, we engineered an E. coli, which is designed to be able to release the NK in the intestine, by inserting the Nattokinase DNA sequence from B. subtilis into pET21a plasmids. In addition, we integrated the BphP1-QPAS1 optogenetic system to manipulate Nattokinase production remotely, along with a detector that could instantly calculate the risks of DVT. To overcome biosafety concerns, we adapted the mazEF toxin-antitoxin system into our kill switch design to mediate cell death once they have been released into the environment.
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HKUST
Title
ShellBi - Prokaryotic Biosensor for Detection of Paralytic Shellfish Toxins

Abstract
Paralytic shellfish toxins (PST), such as Saxitoxin and Tetrodotoxin, are neurotoxins found in shellfish like clams, mussels, and oysters, that cause Paralytic Shellfish Poisoning (PSP) by blocking the sodium channels in neurons. Current methods to test for PSTs suffer from massive drawbacks in terms of ethics, complexity, accessibility, and cost. Hence, we are building a prokaryotic biosensor for the detection of a wide range of PSTs that is ethical, cheap, simple, and direct. The biosensor utilises the antagonistic interaction between Shellfish Toxins and Veratridine to induce significant ionic changes within the cell. An engineered Two Component System in the cells senses this change in intracellular ionic levels and modulates gene expression leading to expression of different reporter proteins. Red fluorescent colour is expected when the toxin is present while green when it is not. Small-scale testing laboratories and even shellfish farms could employ our kit for quality assurance of shellfish.
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WLC-Milwaukee
Title
BlacOps

Abstract
iGEM teams use synthetic biology to solve thousands of problems around the world. From diagnostic tests for cancer to biosensors that detect water contamination, using bacteria and their environment to influence gene expression is commonplace in synthetic biology. However, there are few resources available to those wanting to learn the foundation of all synthetic biology: gene regulation. Especially in urban settings like Milwaukee, a reliable gene regulation curriculum is absent. Furthermore, the concepts integral to understanding gene regulation are intricate, complex, and incredibly difficult to communicate. A reliable, hands-on educational kit simplifies the learning process tremendously. Using the well-documented and understood regulatory components of the lac operon, WLC-Milwaukee created BlacOps, a laboratory kit that effectively teaches the principles of gene regulation.
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Gaston Day School
Title
Risk of Bacteriophage-mediated Lateral Transfer in the Environment

Abstract
Bacteriophages exist in both natural and artificial environments and are able to transfer DNA from one type of bacteria to another. Bacteria, such as E.coli, released from synthetic biology labs may be infected by certain bacteriophages and the antibiotic resistance transferred to other bacteria leading to biosafety issues. To better understand this problem, we will assess the potential risk of lateral transfer of E. coli plasmids by T4 bacteriophages under different conditions. We have shown that T4 can transfer plasmid DNA, which does increase the risk of lateral transfer. However, the number of E. coli phages present in our local environment appears to be quite low, reducing the risk. We added four new T4 early promoters to the registry to support our project. Additionally, our team focuses on providing students who have visual and motor challenges a way to measure liquids through 3-D printing.
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FCB-UANL
Title
Synbiofoam: a synthetic alternative to fluorosurfactants

Abstract
The incidence and intensity of forest and urban fires have increased throughout the last decades; to combat them, firefighting foams containing fluorosurfactants (a group of extremely pollutant chemicals) have been used for years without acknowledging their impact. These have contaminated ecosystems, harming both wildlife and human populations. Thus, we propose the creation of an environmentally friendly firefighting foam that will be produced using synthetic biology techniques. To achieve this, we have continued our work on Synbiofoam, a firefighting foam composed of foaming and stabilizing agents like Ranaspumins (a series of proteins from a native frog) and surfactin and biofilm, both naturally produced by B. subtilis, and whose production will be regulated through a circuit intended to control the bacterium's own regulatory network. As we continue to validate and modify our project based on our stakeholders' responses, we believe that our project will positively impact our society and environment.
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ABSI Kenya
Title
Development of a Portable Biosensor for Detection of Selected Contaminants in Drinking Water

Abstract
Access to safe drinking water is essential for humans. However, drinking water quality is a major source of human infections in low and middle-income countries. Conventional methods for detecting contaminants in drinking water is time-consuming, labour-intensive and expensive. This study aims to overcome this challenge by constructing and validating a cheap, easy to use portable device for the detection of selected contaminants in drinking water. The “Build, Test, Design” cycle will be employed. Appropriate parts will be identified for the selected contaminants and obtained from the iGEM repository. Modelling and simulation will be done to select the best combination. Next, the designed model will be tested for the detection of specific contaminants. Findings from this study will demonstrate the importance of synthetic biology in solving human problems by providing a user-friendly, inexpensive, home-deployable biosensor.
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Stockholm
Title
Mikroskin: an aptamer-based biosensor for the detection of skin microbiome dysbiosis

Abstract
One third of the world's population suffer from skin diseases, like acne and atopic dermatitis. These diseases can cause a negative effect in peoples mental health. There are several sources to skin diseases and dysbiosis in the skin microbiome is one potential source. In order to find the source of the disease, improve to the research community and strive for more personalized health care, we at iGEM Stockholm developed a semi-quantitative rapid test to detect potential dysbiosis that causes the skin diseases. This has been done by developing aptamers targeting teichoic acid that exists in all gram-positive bacteria, which constitutes nearly all bacteria of the skin microbiome. Moreover, other aptamers target C.acnes and S.aureus, which are known to cause skin problems. After conjugating the selected aptamers to 10,12-Pentacosadiynoic acid (PCDA) polymers, a color change visual to the naked eye will be obtained when the aptamers bind to their target.
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NDSU
Title
Anti-fungal recombinant DNA in Escherichia coli for the inhibition of Rhizoctonia solani growth.

Abstract
Crop fungal disease is a global issue that costs billions of dollars in lost crops every year. Crops can be especially susceptible to fungus that infect the root system, like Rhizoctonia solani. Although R. solani threatens many major crops, there are no direct treatments against the pathogen, only management strategies. This project aims to genetically engineer Escherichia coli to produce antifungal compounds which alone, or together in synergy, are effective in inhibiting the growth of R. solani. To accomplish this goal, golden gate cloning was utilized to create several E. coli strains with different antifungal genes. These modified strains were then placed in an antifungal assay with R. solani to establish inhibitory and co-inhibitory effects. Future research would explore inserting these genes in bacteria that have symbiotic relationships with crops to protect against R. solani.
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WVHS SanDiego CA
Title
Project Purified

Abstract
Aridification and rising sea levels are increasing salinity levels and the little-known effects of global warming are drastically affecting coastal plants, like Liquidambar and Jasmine. This project focuses on the HKT1 pathway and overexpresses the pathway in plants battling higher concentrations of salt, to adapt and over time and breed new generations of more-tolerant plants. HKT1-type transporters are key determinants of Na+ and K+ homeostasis under salt stress and they contribute to reducing Na+-specific toxicity in plants. We created four new parts using knowledge from previous literature in which the chassis, E. coli will act as a device to insert a sequence to overexpress the HKT1 gene in plants. The CrGPDH3 promoter gene from C. reinhardtii was found to be inducible under NaCl treatments from previous laboratory experiments, therefore, the project focused on combining the RIA1 section of the CrGPDH3 promoter to initiate overexpression for the HKT1 transporter.
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MIT
Title
B. Syruptilis, a Probiotic Treatment for Maple Syrup Urine Disease

Abstract
Maple syrup urine disease (MSUD) is a rare metabolic disorder characterized by the inability to break down branched-chain amino acids (BCAAs). Untreated, accumulation of toxic byproducts results in irreversible neurological impairment and death. Besides liver transplantation, there is no curative treatment. Those unable to undergo such treatment must strictly control dietary protein intake to prevent metabolic decompensation. However, the gut represents a promising non-invasive intervention point, demonstrated by the amino acid enterorecirculation model. After consulting industry professionals engineering a probiotic treatment for phenylketonuria, a similar metabolic disorder, academic and medical experts, and stakeholders affected by MSUD, we designed a B. subtilis probiotic strain to more efficiently catabolize BCAAs in the gastrointestinal tract. Our approach focuses on overexpressing genes for BCAA importers and catabolic enzymes via a regulatory recombinase switch; and developing an ODE model of probiotic BCAA metabolism based on B. subtilis growth, gene expression, enzyme kinetics, and recombinase activity.
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UNILA LatAm
Title
BioPank - A paratransgenic platform for control of Leishmania transmission

Abstract
Project BioPank is in honor of Pank, a little dog rescued from the streets that had her life shortened by complications from Visceral Leishmaniasis (VL). VL is a neglected tropical zoonosis that affects humans and other animals and has been growing in the region of Foz do Iguaçu and the triple border (Brazil, Paraguay and, Argentina). The SynFronteras team (UNILA_LatAm) will develop a strategy to stop VL transmission by improving the paratransgenesis technique with synthetic biology. We will engineer a Bacillus subtilis to produce an antimicrobial peptide (AMP) inside the midgut of the vector, preventing the parasite from developing, without killing the sandfly. Because of its antibacterial effects, the AMP with leishmanicidal activity DRS-N1, will be produced in its non-activated form and then, activated by enzymes present in the sandfly midgut.
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UGM Indonesia
Title
Auviola: a cyanide-regulating system for gold bioleaching and waste treatment towards sustainable non-mercury gold processing

Abstract
The creation of a sustainable non-mercury gold processing technology has become a considerable challenge. An efficient gold extraction process with less complicated wastewater treatment to minimize the production cost is the ultimate objective of our Auviola project. Bioleaching based on the cyanide-producing bacterium will be engineered, improving the gold extraction and subsequently degrading the off-process cyanide. The Auviola project utilized an engineered Chromobacterium violaceum. To optimize and design the system, kinetic and genomic scale models were demonstrated in both the extraction process and the cyanide degradation afterward. The bioreactor was also designed as the pilot scale for implementing the engineered gold bioleaching process. In the future, this project has the potential to be implemented as a sustainable low-cost gold bioleaching system for the artisanal and small-scale gold miners to bring achievable and beneficial impacts. Moreover, we also contribute to educating the students for understanding and implementing synthetic biology.
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NIT Warangal
Title
Cost-effective enzyme mediated degradation of gossypol in cotton seed meal: A step towards food sustainability

Abstract
Cottonseed is a potential and sustainable source of protein that can satisfy the protein requirement of nearly half a billion people every year. Despite its nutritious and functional food qualities, the consumption of cotton seeds is severely hampered by the presence of a toxic terpenoid known as gossypol. The enzymatic method has proved to be effective for detoxifying gossypol as it is substrate-specific. To date, the degossypolyzation of CSM by enzymatic techniques remains largely unexplored. In the present study, we have computationally screened for enzymes displaying affinity towards gossypol. The top screened enzymes were cloned and expressed in Pichia pastoris and further provided with crude glycerol as a carbon source for cost-effectiveness. Subsequently, the expressed enzymes were verified for their in vitro degossypolyzation ability against CSM. Moreover, the process parameters of enzymatic degradation will be statistically optimized for the maximal reduction in gossypol content of CSM.
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WrightState
Title
De novo Synthesis of α-Terpineol in Escherichia coli Utilizing a Dual Plasmid Model

Abstract
There is a growing demand to reduce production of chemicals via traditional manufacturing. Industries that would benefit from biomanufacturing of compounds via microorganisms include pharmaceuticals, cosmetics/fragrances, and other commercial industries. One molecule with potential applications in these fields is the monoterpenoid α-terpineol. α-Terpineol has been shown to have many medicinal and insecticidal properties. Here, we focused on biomanufacturing α-terpineol as a potential mosquito repellant. Specifically, we aimed to produce α-terpineol via de novo synthesis in Escherichia coli cells utilizing an inducible dual plasmid system. The engineered strain was produced by transforming one plasmid which contained the upstream enzymes of the mevalonate (MEV) pathway and a second plasmid which was constructed to contain the enzymes geranyl pyrophosphate synthase (GPPS) and α-terpineol synthase (αTS). These enzymes convert the end products of the MEV pathway to α-terpineol. Preliminary results show that the system requires further optimization and troubleshooting to result in α-terpineol production.
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NYC B1O
Title
Development of Targeted wild-type Alpha-synuclein Derivatives for Treatment and Prevention of Parkinson's Disease

Abstract
In Parkinson's disease (PD) patients, misfolded alpha-synuclein (aSyn) forms protein inclusions (Lewy bodies) inside neurons, causing neuronal degeneration and symptoms of the disease. Previous studies have shown that increasing the number of chaperon proteins inside the cells or boosting the levels of autophagy, reduces the amounts of misfolded aSyn inclusions. The goal of this project is to target these two processes specifically to the misfolded aSyn instead of indirectly tinkering with cell metabolism. Specifically, the chaperon HSP70 is fused to wild-type aSyn (WT-aSyn). This fusion protein will interact directly with misfolded aSyn converting it into the functional conformation. In the second design, a hydrophobic domain and the LC3-interacting region of SQSTM1 are fused to the WT-aSyn. This fusion protein will drive autophagic membranes to incorporate and degrade misfolded aSyn. Lewy bodies are simulated and monitored using a PD-associated aSyn mutant tagged with the green fluorescent protein.
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UBrawijaya
Title
Clean Harvest of Overexpressed Protein System

Abstract
Indonesia is struggling to independently meet industrial enzyme demands despite efforts to develop cheaper technologies to produce them. Although various enzyme overexpression systems are available, the cost of purification remains a major obstacle. To help address this, we propose the Clean Harvest of Overexpressed Protein (CHOP) system. The system works by anchoring an enzyme of interest (EoI) onto the extracellular terminals of an outer membrane protein (OMP). As the cells grow, they will naturally shed their outer membrane as vesicles (OMV) containing OMP-EoI fusion. Due to OMVs being smaller than cells yet larger than free proteins, they can be separated from the bulk of protein impurities, simplifying the downstream processing. Once separated, the EoI may be released from the OMP by cleaving off a specific linker between the fusion with specific proteases. The development of overexpression systems that simplify downstream purification is expected to significantly reduce enzyme production costs.
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Jianhua
Title
Producing 2'-FL by Saccharomyces cerevisiae

Abstract
2′-Fucosyllactose (2'-FL), one of the most abundant oligosaccharides in human milk, has potential applications in foods due to its infant health benefits. Due to it is prohibitively expensive to obtain 2'-FL directly from human milk, alternative microbial production of 2'-FL is considered promising. Microbial production of 2-FL has been studied mostly in Escherichia coli. To avoid endotoxin contamination and bacteriophage infection in the fermentation process to produce 2'-FL using E. coli, Jianhua aims to produce 2'-FL by engineered Saccharomyces cerevisiae which is generally recognized as safe and has been widely used in food industries. First, using FKP to produce intracellular GDP-l-fucose. Second, Lac12 transports lactose, a fucose acceptor in 2'-FL production into cytosol of S. cerevisiae cells. Finally, FucT2, which catalyzes fucosylation of lactose into 2'-FL using GDP-l-fucose needs to be introduced.
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NYU Abu Dhabi
Title
Rapid detection of fungal pathogens based on electrokinetic concentration and CRISPR-Cas 12a technology.

Abstract
Chytridiomycosis is a fungal disease that has devastated amphibian populations, leading to substantial losses in biodiversity. Caused by the infection of the fungi Batrachochytrium dendrobatidis (Bd) or Batrachochytrium salamandrivorans (Bsal), chytridiomycosis causes rapid decline and extinction of amphibian populations. Both species are responsible for fatal disease; however, Bd infects frogs and toads, while Bsal generally affects salamanders. Amphibians rarely display visible symptoms, limiting diagnosis of chytridiomycosis to post-mortem examination, which highlights the need for diagnostic devices. Current methods for identification of the pathogen are laboratory equipment-dependent, time-consuming, and expensive which contributes to the spread of infection. Thus, our team is developing a diagnostic device that allows for rapid detection of the two disease-causing fungi in a field setting. To bypass the time consuming DNA amplification step, we use electrokinetic concentration of the extracted DNA, followed by a CRISPR-Cas 12a complex binding to the Bd or Bsal target DNA for detection.
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NU Kazakhstan
Title
Remi, du et!

Abstract
Increasing demand for oil & gas production in modern days pushes for higher production and distribution rates. The occurrence of oil spills in such processes is not unusual and the vast majority are left unaccounted for. Existing treatment methods are only efficient for localized spills shortly after the accident, while in the long-term they are less cost-effective and hazardous to Kazakhstani endemic flora & fauna. We propose a solution to this problem by developing a novel agent for crude oil bioremediation - “Remi, du et!”. Specifically, we modify nonvirulent Pseudomonas putida using a dual-inducible system for overexpression of genes nadE and rhlA/B, coding for NAD synthetase and rhamnolipids, respectively. The metabolically-engineered bacteria demonstrate a high yield of rhamnolipids when grown as biofilms under electrofermentative conditions. Since produced rhamnolipids are biosurfactants that emulsify crude oil, the resulting product can be used directly for the treatment of oil spills in ecologically-sensitive areas.
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Ulink-SIP
Title
Efficiently Producing 2′-Fucosyllactose In Escherichia coli

Abstract
2′ -fucosyllactose (2′ -FL), one of the simplest but most abundant oligosaccharides in human milk, has many benefits for the healthy development of newborns. However, the high-cost production and limited availability restrict its widespread use in infant nutrition and further research on its potential functions. This year, Ulink-SIP aims at producing 2'-FL by engineered Escherichia coli. First, we co-expressed the genes for GDP-L-fucose biosynthesis and heterologous α-1,2-fucosyltransferase in E.coli. Then, we deletion genes (wcaJ, nudD, and nudK) involved in the degradation of the precursors GDP-L-fucose and GDP-mannose to further boost the carbon flux from GDP-L-fucose toward 2′-FL synthesis. Finally, preliminary optimization of fermentation conditions was used to achieve a high yield. As a proof of concept, our preliminary 2'-FL production demonstrated a superior production performance.
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Siberia
Title
LEAP2BRAIN

Abstract
The goal of our project is to provide a novel drug delivery system by combining the principles of living therapeutics and N2B delivery. We demonstrate this concept on LEAP2, an endogenous antagonist of the ghrelin receptor GHS-R1a, found in the hypothalamus. The results of an existing mural model suggest that LEAP2 may be used to treat Class I and II obesity in humans.To achieve this, LEAP2 must be periodically supplied directly to the brain. For nasally administered proteins, such delivery is possible via the olfactory and trigeminal nerves, thus bypassing the BBB.We created DH5 alpha and BL21 strains of E.coli that secrete a fusion protein composed of LEAP2, LMWP (a CPP) and a Myc-tag. We're also investigating how CPP's affect membrane permeability to proteins from the "inside". We plan to test our system using an in vitro ALI Transwell system with an artificial mucosal layer.
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GXU-China
Title
Phage vector vaccine for tuberculosis

Abstract
Tuberculosis has been killing millions of people and still cannot be completely defeated. Based on the principle that phage could make bacteria lyse specifically, we proposed to develop a phage vector vaccine for tuberculosis.
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Moscow City
Title
A rapid test system of Equine herpesvirus detection based on CRISPR/dСas13

Abstract
Equine herpesvirus outbreaks lead to mass deaths and quarantines. Disease symptoms can be easily confused with other diseases. Current diagnosis methods - PCR-tests - are time-consuming. Therefore, we decided to create a test system based on CRISPR-Cas. It detects herpesvirus by its RNA. We use deadCas13 protein, connected by a linker with two halves of reporter protein - beta-lactamase. Two deadCas13 proteins are directed by gRNA to the RNA of the virus. When the distance between two Cas-proteins is correctly selected, two beta-lactamase proteins can fuse into a functional protein that destroys nitrocefin — the solution's color changes from yellow to red. To avoid false-positive results, we use two guide RNAs. With one guide, RNA reporter protein will not become functional. We are willing to create a system that will not require any special skills to use it. Horse owners will be able to detect herpesvirus using only saliva.
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ACIBADEM ISTANBUL
Title
Rapid Easy Point of Care Diagnostics by Combining Antigen Tests and Nucleic Acid Amplification

Abstract
Nucleic acid amplification tests like polymerase chain reaction have high sensitivity and are the standard test for the diagnosis of infections like Covid19. However, they require molecular biology laboratories and trained personnel. Therefore, they are not point of care (POC) diagnostics. Rapid antigen tests fix an antibody to a chromatic paper strip, detecting antigens binding to this antibody. However, since there is no amplification, sensitivity is low. Combining the principles of these methods, our kit works by binding a molecule of the infectious agent on a strip via antibody or DNA probe, then doing signal amplification on the bound molecule. This involves an innovative isothermal nucleic acid amplification at room temperature, not requiring sophisticated instruments. The nucleic acid amplification depends on a synthetic original enzyme that can cut one strand enabling strand displacement by DNA polymerase. This method may enable rapid diagnosis of diseases at POC and help control pandemics.
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SDSZ China
Title
City Band-aid

Abstract
Building repair is crucial when it comes to ancient buildings, since cracking impacts hugely on buildings' durability (one of the most important features to be considered during construction). Our team, SDSZ_China, uses a new microbial technology, which has the advantages of green environmental protection as well as real-time remediation, to help fix this issue with its great potential for development.
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USP-EEL-Brazil
Title
Honorato

Abstract
Ophidic accidents are categorized as a neglected tropical disease by the World's HealthOrganization (WHO), which means that even though there is a high incidence of theseaccidents, they don't get enough attention.In Brazil, Bothrops is responsible for 85% of snake bites, with 20,6% resulting in necrosis,therefore our project is focused on producing a recombinant γPLI, a broad spectrum inhibitorfor the treatment of ofidic accidents with the Bothrops genus. Our inhibitor aims to actagainst phospholipase A2, one of the main groups of enzymes present in the venom,responsible mainly for the necrosis that also has a synergyc effect with the other enzymes.The expectatives with our project is to provide to our community a faster and more accessibletreatment, hence, reducing the sequels and death rate due to Bothrops ophidic accidents.
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Tec-Monterrey
Title
Diagnosgene

Abstract
Agave is a crucial part of Mexico's culture, ethnicity, and economy. In our country, nearly 558 municipalities are dedicated to its production, and its use for the manufacture of tequila generates a net income of more than 1 billion USD every year, but its production faces an obstacle: the wilting of agave, which is caused by a diversity of fungi and leads to the eventual death of the plant.Current detection systems are either visual or laboratory examinations, but the process can take up to three weeks, and by that time the infection can be spread through the whole land. Our proposal, Diagnosgene, consists of a rapid, easy-to-use, early detection system which can be implemented on the field. The detection which occurs within a cell-free system, is accomplished by the usage of toehold switches, which are RNA tools that get specifically attached to a given complementary sequence.
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Shanghai City United
Title
Innovative poultry beverage that contains lactobacillus that produces xylanase

Abstract
Non-starch polysaccharides (NSP), such as xylans, are one of the most abundant substances found in grains but few animals can digest them well. We noticed Xylanase, a type of enzyme that degrades the linear polysaccharide xylan, which could be served as a dietary supplement to treat the poor digestion issue. In this project, we introduced a probiotic which was previously genetically edited by us to possess the ability to secret Xylanase and form a feed that can be applied in monogastric's, mainly poultry's daily meals. The product will be designed as a type of “beverage” to make up for the poultry's insufficiency of the xylan digestive enzymes In this case, poultry will be able to absorb more energy when given the same amount of feed, which will further ease the food resource shortage in the world.
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UCDavis
Title
UC Davis iGEM 2021 Project Cargo

Abstract
Team UC Davis seeks to optimize mRNA delivery in vitro for the foundational advancement of mRNA vaccine technology, namely to reduce side effects caused by an innate immune response due to sudden influx of spike protein. We plan to insert an RNA secondary structure known as an Iron Response element (IRE) into the 5' UTR that reduces the sudden influx of spike protein by gradually releasing the available mRNA transcripts for translation. The protein known as an iron response binding protein (IRBP1/IRBP2) binds and blocks translation until an iron allosterically binds and IRBP/1IRBP2 unbinds to allow for translation. A working pipeline was created to automate the insertion process and structural stability by utilizing the RNA folding package, Vienna. To prove the mRNA/IREBP system constructs were expressed transiently in mammalian cell cultures and translation was monitored with destabilized YFP. The design is modular and able to integrate into any mRNA therapy.
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Shanghai Metro HS
Title
Ceres: Silage additive for the gastrointestinal health of livestocks

Abstract
Our project aims to create a gene-edited E.coli bacteria as a silage additive that can help improve the digestion of ruminants on silage and further improve the quality of the livestock. In the selection of the desired gene, PKC-001 from Pseudomonas aeruginosa was reported to secrete the cellulase which maintains well activity under both acid and base conditions. Therefore, we optimized its codon and further constructed the plasmid to combine the optimized gene-PKC-OP, which was transformed into the E. coli for expressing itself. Even though we encountered many difficulties in lab work, we finally obtained our engineered E. coli which could express the desired cellulase. We hope this silage additive could contribute the stockbreeding development and improve life quality in the future.
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Shanghai HS ID
Title
Applications of CRISPR-Cas 9: Increasing the Expression Efficiency of Foreign DNA in Lactobacillus casei

Abstract
Lactobacillus casei was found to have the potential to cure and prevent common health issues like high cholesterol, hypertension, and gastrointestinal diseases. Recent studies show that L. casei's capabilities in medicine and food are highly restricted by their low transformation efficiency of foreign DNA, due to its restriction-modification system. Our purpose is to improve the transformation efficiency of foreign DNA in L. casei to maximize their benefits. In the laboratory, we constructed a plasmid containing CRISPR-Cas9 in E.coli, then transformed the plasmid into L.casei so that the target gene LSEI-2094 can be removed and the restriction-modification system can be silenced. Lastly, we confirmed the results through both sequencing and controlled experiments to verify the efficiency before and after the modification. Results show that the transformation efficiency has significantly increased post-modification, it is hopeful that this modified L. casei could be applicable in pharmaceutical and food.
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CUHKSZ
Title
EthaNO

Abstract
Alcoholic beverages usually play an important role in our social life. Some spirits, like Chinese Maotai, may also contain some ingredients beneficial to human health. However, the high concentration of ethanol in those spirits can also take a serious toll on human health. Our project EthaNO is a tiny device that can decompose ethanol in alcoholic beverages. We aim to figure out the oxidation mechanism of bi-functional alcohol/aldehyde dehydrogenase (AdhE) through computational biology E.coli bacteria are transformed with modified gene and will generate enzymes to degrade ethanol. To make it user-friendly, PVA-SA granules are applied to store the bacteria in the device. We will optimize the immobilization method to maximize degradation efficiency. With EthaNO, people can enjoy alcoholic beverages like Maotai without worrying about ethanol.
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Bonn-Rheinbach
Title
BioLan – Protein-based extraction of rare earth elements from ores

Abstract
Lanthanides form the majority of the highly important rare earth elements (REE) and possess outstanding catalytic and magnetic properties. REEs are essential in all fields of modern technology – from automobility to medical diagnostics. The current REE mining is energy-intense and relies on many synthesized chemicals.We developed a protein-based process to increase sustainability and selectivity of REE mining. The bacterial protein lanmodulin (LanM) was heterologously produced in E. coli and purified in a novel, chromatography-free one-step method. LanM was added to an ore leachate to form an REE-LanM complex. After isolation of this complex, REE ions were released by pH reduction. Upon removal of LanM, this biological chelator was regenerated and a highly pure REE solution was obtained. Our project is characterized by iterative process optimization tailoring the approach to industrial application.Overall, it was demonstrated that the protein LanM can replace current chemical chelating agents in REE mining.
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Open Science Global
Title
Frugal Biofoundries for Distributed Enzyme Manufacturing

Abstract
Biotechnology can help avert human civilization's looming mass extinction caused by fossil energy. However, intellectual property, skills, and high costs of hardware and wetware create massive barriers. These barriers were profoundly felt throughout the pandemic as sparse access to bio-manufacturing infrastructure severely limited medical relief, testing, and vaccination. With an internationally distributed team of 10 nationalities and 4 laboratories, we aim to break these barriers by frugally producing and purifying thermostable enzymes essential for synthetic biology solutions. We have established a pipeline for enzyme production in frugal bio-foundries that's based on open-sourced software, hardware, material transfer agreement, and wetware constructs of B. subtilis for reducing protein extraction costs. Our genetic constructs will be freely distributed by FreeGenes. To gauge the global needs of implementing a frugal bio-foundry in different parts of the world, we set up a hackathon in partnership with JOGL. Our project contributes towards democratizing biotechnology.
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Yonsei Korea
Title
Detection of rice blast fungus Magnaporthe oryzae using Cu2+ dependent DNAzymes and gold nanoparticles

Abstract
Rice blast, a fungal disease caused by Magnaporthe oryzae, is increasingly affecting rice plants leading to approximately 30% of rice production losses globally (Nailey 2016) and 15-60% annual incidents in Korea. The current treatment and diagnostic methods, however, are not environmentally friendly or are not easy to use for farmers. Therefore, we propose a DNAzyme based detection system to specifically detect the mif23 gene of M. oryzae. By Cu2+ catalyzation, our DNAzyme cleaves at the ends of the CTGC site of the target gene. We used PAGE and denaturing PAGE to validate the specific cleavage activity. We also confirmed that Cu2+ is significant in catalyzation, and that it is an efficient system in detecting the target gene. Lastly, gold nanoparticles functionalized with the complementary target DNA sequences to the cleaved DNA products were employed as a colorimetric approach to visualize the DNAzyme catalyzed cleavage reaction.
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まとめ


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大変多くのチームが、オリジナリティ溢れたプロジェクトを行っていることがわかっていただけたかと思います。本記事では、iGEM2021に出場したチームの1/4ほどのチームしかとりあげられておりません。さらに興味をもっていただいた方は、その他のバージョンも参考にしていただけたらと思います。


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