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Collaboration

National University of Singapore

Life Hacks! 2.0

Life Hacks! 2.0 is a one-day outreach event organised in collaboration with Team NUS_Singapore to educate pre-tertiary students about synthetic biology and bioentrepreneurship. More than 70 junior college and polytechnic students participated in the fun-filled event. The event lineup includes a series of lectures by professors from local universities about how synthetic biology is solving real-world problems, a panel discussion between seasoned bioentrepreneurs, interactive games designed to expose students to various concepts and topics in synthetic biology and an iGEM project sharing session by both teams. We also hosted the team leader of Nazarbayev University iGEM 2018, Daulet Aitymbayev, who gave a great talk about their previous iGEM project on engineering cyanobacteria for oil refining!

Group photo with Team NUS!

Meeting Team Macquarie for the first time!

Macquarie University, Australia

Wet lab collaboration

In collaboration with Team Macquarie, we performed qPCR measurements for 4 of their constructs to verify riboswitch expression upon IPTG induction at different time points (0h, 4h and 24h). In exchange, they performed SDS-PAGE on nuclear pellets collected from our transfected cells to verify that RNA editing activity of our constructs correlates to its nuclear localisation. Although the presence of editing from our data provides evidence that our constructs work as intended, their additional experiments on protein expression has added a layer of assurance to our project. The transfected constructs were H460D (Rx-hsADAR 4) and H460A (Rx-hsADAR 5). We also sent 2 plasmids (dCas13b and dCas13d) for them to help us check its protein expression in E. coli for the possibility of purification.

Figure 1A shows the presence of our dCasRx-ADAR2_DD constructs (~160kDa) in the nuclear pellet of our transfected cells. The bands may appear faint due to degradation during transport, however, this indicates that our constructs were localised in the nucleus for editing as intended by the NLS tag. We were also interested in testing the applications of dCas13b and dCasRx in vitro. As we did not have access to equipment for protein work, we gave Macquarie both plasmids to test their expression in E. coli for possible purification in the future. From the SDS-PAGE (Figure 1B), it seems that only dCas13b was able to be expressed but not dCasRx. This may indicate that we should add a solubility-tag in the future. The collaboration was a fruitful one, with both teams gaining insights into their projects. This was also the 4th year of our collaboration and we hope to continue this tradition in the future. We wish Macquarie all the best!

Figure 1. SDS-PAGE results. (A) Presence of our constructs in the nuclear pellet of HEK293FT transfected cells. NP: nuclear pellet. (B) Protein expression of dCas13b and dCasRx.

University of Düsseldorf, Germany

International postcard exchange

We were excited to be part of Team Düsseldorf's international postcard exchange which showcased the projects by different iGEM teams and fun memes about synthetic biology. From these postcards, we learnt about what international iGEM teams were up to and also got to share about our Cas13-ADAR RNA editing system as well.

Unboxing the postcards we received from iGEM teams all over the world!