Team:NUS Singapore-A/shadow/Collaborations
1. CUHK
NUSGEM initially wanted to explore using RNA aptamers as a reporter system for our stress promoter (htpG1) and blue light repressible promoter. However, we faced a lot difficulties in constructing our aptamers reporter system and we could not get a successful construct for many months. However, we read that the CUHK iGEM team was working on aptamers. Hence our team leader, Nanda, met up with them in Hong Kong.
After an exciting discussion at the Lee Woo Sing College of Chinese University of Hong Kong, both teams saw a great collaboration opportunity, discovering many similar aspects of our projects in wetlab, hardware and modelling.
The two teams concluded our meeting by agreeing to keep in contact, and we continued to discuss intensively on our collaboration even after Nanda returned to Singapore.
As the CUHK team was working on a RNA-based project, they were urgently looking for a particular strain of E. coli, BL21 Star. It is a RNase-knockout strain to maximize RNA production. They asked whether our lab has it - we do! Without hesitation, we shipped the strain over to them within a week.
Because NUSGEM needed help on RNA aptamers constructions, Team CUHK offered their expertise by redesigning our RNA aptamer constructs and designing suitable primers to increase the chances of successful construction. In addition, they incorporated the RNA aptamer that they aimed to characterise, iSpinach, with our stress promoter (BBa_K2819010) for us to characterise. Both teams agreed on constructing the same constructs, including those that they hoped for us to characterise, using the same set of primers. They advised us to use overlap PCR in our constructions, a technique that we don’t usually use in our lab. We took their advice and our gel electrophoresis results below show that our overlap PCR was successful!
With their help, we were finally able to successfully construct the RNA aptamer plasmids! After successful constructions of the aptamer constructs, NUSGEM assisted CUHK in characterising them. We measured the fluorescence intensity given off by the aptamers under different temperatures, and found that that lpp-tRNA-iSpinach gives off a higher fluorescence reading than lpp-tRNA-Spinach 2.1 (Figure X)!
(Figure showing characterization results - get from CUHK?)
CUHK also hoped to carry out their survey with a wider audience, so NUSGEM not only helped them in their survey, but also spreaded the word of their survey to help them to gather responses in Singapore.
2.Toulouse and Quebec
What started out as a contacting session between Toulouse and Quebec turned into a great collaboration between three groups from different parts of the world. All three teams wanted to bring to discussion how synthetic biology touches different aspects of . The three main subjects we chose to discuss were bioethics in synthetic biology, economic aspects and lastly, society’s perspective of synthetic biology.
3.NTU
With Team NTU just being next door in sunny Singapore, how could we not collaborate with them? Both teams first met in our SG iGEM Meetup (hyperlink), and we subsequently met up several times to finalise our collaboration plans.
NUSGEM help Team NTU to verify and characterise their dCas9 fused with adenine base editor system in E. Coli BW25141. We co-transformed 2 plasmids into the strain, ___ and ___, and characterised the constructs under 4 different inducer conditions: No inducer, IPTG, Arabinose, IPTG+Arabinose. Their system has the following proposed working mechanism: Kan*(to confirm the name), induced by IPTG, is a construct with a mutated kanamycin gene attached to SgRNA, while dCas9 (to confirm the name), induced by arabinose, is capable of editing and correcting the mutation on Kan* to restore the plasmid’s antibiotic (kanamycin) resistance.
The editor system is only supposed to work when both IPTG and Arabinose are present, as both SgRNA and dCase9 must be induced. SgRNA provides a guide sequence to direct dCas9 to target its plasmid, to perform adenine editing on Kan* (to cfm plasmid name). To demonstrate this, we plated the cells from each condition into 2 types of agar plates with different antibiotics composition, one with kanamycin+gentamycin+chloramphenicol, and the other with gentamycin+chloramphenicol.
Top row: Agar plates with gentamycin+chloramphenicol (control)
Bottom row: kanamycin+gentamycin+chloramphenicol
Left to right: No inducer, IPTG only, Arabinose only, IPTG + Arabinose
We verified that their system works as proposed. As shown in the picture above, only the system induced by both inducers had colonies growing on the agar plate with kanamycin+gentamycin+chloramphenicol, while no colonies were observed for the other 3 induction conditions.
As our labs lack the necessary equipment to conduct RT-qPCR, we were unable to study the level of mRNA expression of our constructs. Fortunately for us, Team NTU is well-versed in this technique, having provided assistance to many teams previously. We gratefully accepted their offer to help and passed them several of our dye-synthesizing constructs for RT-qPCR analysis and characterization. The results can be seen in Figure X.
The low levels of expression helped to pinpoint the reason for the lack of success we have in producing our target compound. It prompted us to make revisions to several aspects of our project, most notably the change of host from BL21 (DE3) to BL21 Star (DE3), a RNase knockout strain which has enhanced protein expression. Moreover, we also gained further insights into the design of our plasmids and how we can improve on them.
4.Elaboration
Michigan State, Newcastle, Uppsala, TUDelff
Other potential collaborations were discussed over Skype sessions with (list). We shared and discussed ideas about genetic circuit designs, human practices, fund-raising. We were happy to chat and make friends all over the world, and we can’t wait to see you guys at the Giant Jamboree!
Click any slide for more
[8/8] - Dr. Yvonne Chow
In Progress
[8/8] - Dr. Yvonne Chow
In Progress
[8/8] - Dr. Yvonne Chow
In Progress
Visit from Jiangnan iGEM team
On 18 July 2018, NUSGEM was privileged to host 2 members from the Jiangnan iGEM team!
The Jiangnan iGEM team members were in Singapore for a summer exchange programme, and they chanced upon our project. They took the initiative and wrote to us about the possibility of a meet-up to share our respective team’s iGEM experience, and learn more about the problems we have encountered, and how we managed to overcome them.
Without hesitation, we agreed to meet up with the Jiangnan iGEM team members! In fact, we were all so excited about the meet-up that we planned an engaging day of activities for our fellow iGEM-ers!
Firstly, we invited them to the BioMakerSpace, which is where our NUSGEM team carries out most of our experiments. At the BioMakerSpace, we brought them on a lab tour, which culminated in a fruitful sharing session on our projects, where we even exchanged solutions on the problems we faced. After the laboratory tour, we brought them on a NUS campus tour, visiting the various libraries (there are more than 6!) in our school, as well as the biofoundry in SynCTI (Synthetic Biology for Clinical and Technological Innovation).
The two teams had a great time interacting with each other and we can’t wait to see each other again in the Jamboree!
[8/8] - Dr. Yvonne Chow
In Progress
[8/8] - Dr. Yvonne Chow
In Progress
[8/8] - Dr. Yvonne Chow
In Progress
SGiGEM Meet
On 11 June 2018, all three Singaporean iGEM teams met up at the National University of Singapore, for the first ever Singapore iGEM meet!
The goal of our inaugural meet-up was to provide an opportunity for all Singaporean iGEM participants to get to know each other, network, and spark fruitful discussion on potential collaborations. This meeting was the first official platform for all three teams (NUS_Singapore-Sci, NUS_Singapore-A and NTU-Singapore) to share with each other our respective team projects, and discuss possible avenues for collaboration. Hearteningly, members from all three teams felt the meet-up was fruitful, and we made many exciting plans for collaborations!
No meeting is complete without food! All of us had a great time socialising and discussing our iGEM experience over a pizza party! The meetup was indeed a great opportunity to get to know members from other teams, and even our own team members better, and perhaps more importantly, many memories were made that night!
HEAR OUR STORY
We’re using synthetic biology to change the way the world thinks and functions.
We’re a little disruptive, dangerous and maybe a tad too bold. But the world needs some shaking up every now and then, and we think we’re just the right people for that.
Together we’re going to engineer biology and create something awesome.
CONTACT US
nusigem@gmail.com21 Lower Kent Ridge Rd, Singapore 119077