Please browse our collection of projects and their respective iGEM wiki sites. We are proud to work with a determined group of students each summer and to have strong support from faculty advisors, and internal/external funding to make these projects possible.
PHARMING THE BLUES
BREAKING THE ICE
CHIMERIQ X SYNTHETIQ
Breaking the Ice: Improving Antifreeze Proteins for Practical Use
Each year, increasing numbers of individuals are added to organ wait lists worldwide. This is met with an ongoing shortage of donor organs, in part, limited by preservation technologies. A human heart can currently be stored for 6 hours before significant tissue damage results in a non-viable organ. QGEM aims to use antifreeze proteins (AFPs), natural proteins that enable certain organisms to survive in sub-zero climates, to rectify this limitation. We have engineered two classes of AFPs focused on improving protein function and stability, respectively. Our primary objective has been the development of an anchoring system to increase local concentration of AFPs by attachment to a self-assembling scaffold. This system increases the probability of favorable interaction with ice surfaces thereby improving AFP activity. Our secondary project structurally modifies an AFP, enabling the protein to withstand a more diverse chemical environment. This increases potential industrial applications in food and energy sectors.
Found across all domains of life, inteins are protein elements capable of autocatalytic splicing. This phenomenon, termed protein splicing, allows inteins to be a point of control in forming mature and functional protein products. Current genetic circuits are limited by transcriptional lag, rendering them limited in applications which require fidelity and responsiveness. Inteins represent an innovative solution to improving the sensitivity of biosensors and synthetic circuits circumventing this lag and controlling the output of the circuit at the protein level. Here, we have created an intein toolkit, designed to introduce and facilitate the use of inteins in future iGEM projects. We have engineered several inducible intein switches capable of being activated by common environmental triggers as well as small molecules. To showcase the utility of inteins, we have also developed an intein based system to tackle mitochondria disease by allotropically expressing and transporting mitochondria proteins using inteins.
Biosynthesis and breakdown of human odour compounds for the behavioural manipulation of malarial mosquitos
Malarial mosquitoes are developing resistance to key insecticides and drugs, and are becoming diurnal to avoid treated mosquito nets. Recent studies have shown that the African mosquito uses human foot odour to locate its host, a trait that is enhanced when the insect is carrying malaria. We plan to combine a carboxylic acid reductase and an acetyl transferase, in order to create E. coli capable of converting a major component of foot odour (isovaleric acid) into banana smell (isoamyl ester). This could have both commercial and humanitarian applications. Our second goal is to deliberately synthesize mosquito attractants inside traps. Recent research has shown that a mixture of CO2 and foot odour volatiles can be more attractive than a human. We have chosen indole as our first attractant, a compound naturally found in human sweat. We hope our project will show that bioremediation and biosynthesis techniques have applications in mosquito control.
ChimeriQ x SynthetiQ: Chimeric flagella scaffold enhancing bioremediation and manufacturing, presented with dance!
This year, Queen’s iGEM team is using flagella to host heterologous proteins that will result in thousands of useful enzymes organized in an extensive scaffold, with the benefits of extracellular synthesis, degradation and arrangement. The fliC (flagellin) protein is known to spontaneously polymerizes to form the length of flagella in E.coli. By replacing the variable D3 domain of the fliC protein with proteins for binding, degradation, adhesion, and synthesis, we can increase the efficiency of bioremediation and biosynthesis, and facilitate the collection of products in situ or ex situ. This year we will also introduce dance as a presentation form and part of our human practices project. Known as SynthetiQ, we will be the first group ever to use dance to replace powerpoint slides at a research conference.
Nemoremediation: Engineering C. Elegans into a Toolkit for Soil Bioremediation
Naphthalene is a pollutant produced by oil sands operations. The Queen's team has engineered the nematode worm C. elegans into a toolkit for dealing with this compound in the soil. We have produced constructs with GPCRs from M. musculus, R. norvegicus, and H. sapiens intended to enhance the worm's ability to chemotax toward naphthalene. We are working on a field bioassay based on fluorescent proteins that will indicate the presence of naphthalene in a soil sample. The goal is to have a population of green fluorescent worms chemotaxing toward and a population of red fluorescent worms chemotaxing away from the napthalene in the soil sample. Finally, we have biobricked the P. putida gene nahD, which encodes a degradative enzyme as part of a naphthalene catabolic pathway. The nahD gene encodes the enzyme 2-hydroxychromene-2-carboxylic acid isomerase, which catalyzes the fourth step in the catabolic pathway.
WormWorks: Introducing the nematode C. elegans as a multicellular chassis
Engineering C. elegans for a powerful iGEM toolkit for multicellular organism projects
Historically, the iGEM competition has tended away from working with eukaryotic and multicellular organisms, limiting prospects for higher levels of project complexity in favor of simpler and easier-to-understand bacteria. The nematode worm Caenorhabditis elegans was examined as a prospective chassis for use in the competition. Once it was decided that the opportunities presented by the organism appeared to outweigh the challenges involved in working with it, a foundational library of parts was built and tested within the organism. This collection includes useful promoters, reporters, effectors, and a terminator. An educational resource specifically targeted at iGEM participants was written and incorporated into the team wiki in order to assist future teams in learning about and exploring the possibilities offered by C. elegans.
All living organisms contain ribosomes, molecular machines made up of proteins and rRNA that perform translation of messenger RNA (mRNA) to protein, within their cells. More recently, cellular machinery that synthesize proteins without the use of ribosomes, termed nonribosomal peptide synthetases (NRPS), have been discovered in some bacterial and fungal species. These enzymes consist of a chain of modules, each module contributing a specific amino acid and modifying the growing peptide as it is passed along the chain. In this manner, a unique peptide can be synthesized without the traditional ribosomal unit or mRNA. The genes necessary to create one specific NRPS peptide are usually found in close proximity to each other in the genome as an operon or gene cluster.
Glacial Gladiators: Bifunctional Biofilms for Arctic Bioremediation
Biofilms are often maligned because of their roles in antibiotic-resistant infections and dental plaque. However, biofilms also offer an attractive platform for the design of self-assembling biomaterials programmed for specific functionality. The amyloid protein CsgA accounts for the majority of the proteinaceous component, curli nanofibres, of Escherichia coli biofilms. CsgA has been shown to be tolerant of C-terminal fusions, allowing CsgA endowed with diverse peptide domains to be secreted and self-assembled extracellularly similar to normal curli nanofibres. We present the genetic engineering of CsgA to create a biofilm that binds ice and degrades hydrocarbons. A type I antifreeze protein, AFP8, will be fused to CsgA for ice binding, and a PA-14 adhesin domain will be fused, via the SpyTag-SpyCatcher system, to bind the hydrocarbon-degrading bacterium Marinobacter hydrocarbonoclasticus. Thus, the end-product will be a bifunctional biofilm capable of establishing itself on Arctic ice to degrade toxic hydrocarbons present in oil spills.