Team:Macquarie Australia/Collaborations
Singapore iGEM Team Collaboration
Revisiting our successful collaboration with the NTU Singapore team last year, we approached them to see if this year’s team would be interested in partnering up again. After discussing our respective projects, a joint decision was made on how to best help each other.
Knowing that we have expertise in protein purification and in vitro assays, NTU incorporated His-tags into the constructs that they wanted to test and requested for us to purify and test their dCas9 constructs in vitro.
In return, knowing that the NTU team is experienced in quantitative PCR from our collaboration last year, our team was interested to know the different transcription levels of the genes within our functioning Hydrogen Gas Producing Gene Cluster plasmid this year (HGPGC) as it contains a few lac promoters within the gene cluster. We asked NTU Singapore to perform RT-qPCR after induction using IPTG to study the expression of our proteins in the gene cluster.
Our instructor delivered our HGPGC plasmid to perform RT-qPCR to the NTU team during his holiday in Singapore, and brought back the plasmids containing the clones that they were interested for us to test on his return trip.
The in vitro assay was performed on four truncated dcas9 mutants: wild type dCas9 (WT), 3P, 5P and HNH. This project used the dCas9 enzyme to form a Ribonucleoprotein (RNP) complex with Ferrochelatase (the target sequence tested). The results from the Electrophoretic mobility shift assay (EMSA) show that the dcas9 mutants (WT,3P, 5P and HNH) were successfully truncated, and no longer retain their nuclease activity (see Figs. 1-3). This is evident from the gel shift present, the shift of the target sequence protein and the presence of gRNA, indicating the mutant dCas9 has bound the target sequence of DNA from its slower migration through the gel (see Figs. 1-2).
Summary:
- Through their RT-PCR expertise, NTU Singapore aided us in gaining a better understanding of the transcription rates within our composite Hydrogen Gas Producing Gene Cluster plasmid.
- Our efforts have successfully purified their dCas9 mutants and suggest that all are functional in vitro. This has provided in vitro evidence for the NTU Singapore team that these four mutants are no longer functional nucleases, but retained their ability to bind the gRNA and targeted gene sequence.
Manchester Collaboration
H2ydrogem collaborated with iGEM teams from nine other countries (EU, USA, Brazil, Japan, Chile, Indonesia, Korea, Canada and India) to create a comprehensive document on GMM (Genetically Modified Materials) legislation around the world. Each team conducted extensive research into the laws surrounding GMM inclusion and labelling in commercial products. This research was then collected and summarised to produce integrated responses to the following six questions:
- 1. What institutional body enforces the laws regarding the use of GMMs?
- 2. What institutional body enforces the laws regarding the use of GMMs?
- 3. Who regulates the use of GMMs on a case-by-case basis?
- 4. What legal requirements do I have to fulfil in order to be able to use GMMs in my business?
- 5. As an employer who exposes their workers to biological agents, what safety measures am I obliged to adopt?
- 6. As a consumer, how do I know if a product I want to buy contains GMMs?
- 7. What are the most important documents related to GMMs?
This document can be viewed on the Manchester iGEM team’s page. We used this document to assist in the formulation of our business plan (particularly our distribution strategy) and our safety protocols.
Other Collaborations
Pedestal
With help from Mr Michael Rampe and Mr Peter Reeves from Pedestal our team designed a 3D rendering of our prototype, H2GEM, which will soon be available on their website. With this rendering, our team was able to conceptualise our design in the physical space for its potential use in marketing H2GEM.
The Australasian iGEM Social
During the proceedings of the Synthetic Biology Australasia Conference, our team met up with our colleagues from Melbourne, Auckland and University of Sydney in Kirribilli, and presented our projects together during the iGEM session of the conference. The two days of the conference were capped off with social drinks and an impromptu trip to Luna Park, which was kindly organised by our iGEM Representative, Abigail Sison.
Team Franconia Game Contribution
“There has been an incident at the iGEM Giant Jamboree 2017 in Boston and numerous young scientists have been contaminated with some of the most dangerous viruses of our time!” Team Franconia developed an app that will run an augmented reality game, for which the Macquarie Australia iGEM team, H2ydroGEM, will contribute by attaching a small QR-code (approx. 4 by 4 centimeters/1.5 by 1.5 inches) on our station, for players to partake in the game, as well as attract our iGEM family to our station.
Rate My Gel
H2ydroGEM is contributing to this game developed by the team from the University of Sydney, adding to the thrill of the iGEM spirit.
Methane Emissions
The H2ydroGEM team helped the 2017 UNebraska-Lincoln team in providing an Australian perspective to the issue of methane production.
Ainsley Newson (A/Prof of Bioethics at Sydney Health Ethics, Sydney School of Public Health)
Guided us in ensuring our team kept in line with various ethical concerns involved.
GOLD SPONSORS |
BRONZE SPONSORS |
LOCATIONMacquarie University Balaclava Road, North Ryde, NSW, 2109, Australia E7B 350 CONTACT US |
FOLLOW US ON
|
