Team:CityU HK/Collaborations

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The Hong Kong University of Science and Technology iGEM Team - ShellBi

Meeting with the HKUST iGEM Team

We met with the Hong Kong University of Science and Technology (HKUST) iGEM Team – ShellBi – for the first meeting on 16th July 2021 via zoom. We presented and shared details of our projects at the zoom meeting. ShellBi comprises a diverse team where students are from different areas of study and are of various nationalities. The HKUST iGEM project addresses a local problem on paralytic shellfish poisoning (PSP) which adversely affects the Hong Kong aquaculture industry. The current testing methods are inhumane and take too long to provide results. As such, they proposed to develop a biosensor based on an E. coli two-component system to detect osmolarity changes (measured by fluorescence intensity) that result from the antagonistic action between the two compounds – Saxitoxin (STX) and Veratridine (VTD).

Figure 1. Meeting with HKUST iGEM Team (on 16th July)

In the meeting, we discussed our projects further and agreed to conduct project opinion surveys on our respective projects in both of our universities. Additionally, they discussed with us their plan to organize a Hong Kong iGEM Symposium for this year and invited our team to present our iGEM project at the event.

Hong Kong iGEM Joint Symposium 2021

The 2021 Hong Kong iGEM Joint Symposium on “Biotechnology is the Future” was held on 11th August 2021 and was primarily organised by the HKUST iGEM team with presentations given by several of the iGEM Teams in Hong Kong – CityU, HKU, United Christian College, Hong Kong International School, among others. We were delighted to participate in the event, and we treasured the opportunity to share our project with the other Hong Kong teams as well as learn more from their projects.

We were also fortunate enough to learn about Prof. Becki Kuang’s research on prolonging the half-life of synthetic mRNA in her talk, and how it has the potential to revolutionize vaccines. She is recently focusing on the development of cell fate controlling RNA machinery and high-performance mRNA drugs. A particularly interesting part of her talk was the significance of the Poly(A) tail of an mRNA strand in controlling its half-life and how her team has discovered modifications that can be used to extend the protein expression duration of an mRNA strand to over 48 hours. Her team believed that modified poly(A) tail can greatly impact mRNA therapeutics by reducing the number of mRNA administration required per treatment and for minimizing the dosage of mRNA therapeutics.

During the mingling session after the talks, we communicated with every team at the 1-day Symposium and exchanged feedback on each other’s projects for further improvement. We also recorded a vlog of our entire day’s experience to provide the public with an insight and better understanding about Synthetic Biology as well as the topics discussed in the Symposium. The talks presented by all speakers at the Symposium and the conversations with other iGEM teams undoubtedly broadened our knowledge about the scope and capabilities of synthetic biology.

Figure 2. Presentation by the CityU iGEM team at the HK iGEM Symposium (on 16 Aug 2021)

Figure 3. HK iGEM Joint Symposium (on 16 Aug 2021)

The University of Hong Kong iGEM Team – Evolve

Meeting with the iGEM Team of the University of Hong Kong

On 30th June 2021, we met with the University of Hong Kong iGEM Team – EVOLVE. Although both iGEM teams met through zoom for the first time, it did not take long to break the ice to start a fruitful and interesting conversation. After mutual introduction of our teams, we presented each other’s iGEM projects for this year’s competition. The HKU Team was aiming to tackle the problem on plastic pollution and proposed to bioengineer an “E. coli – Cyanobacteria” plastic degradation system, which is similar to what our 2020 CityU iGEM Team had worked on.

We also agreed to conduct project opinion surveys on our respective team projects through our university networks for better outreach and collection of more comprehensive data for analysis. We managed to receive a good amount of responses from the HKU iGEM team, which aided our survey hitting 142 responses.

Figure 4. CityU-HKU Online Meeting (on 30th June)

Preparing the PETase Plasmid for the HKU iGEM Team

Since, the HKU 2021 iGEM Project is similar to the previous year’s CityU iGEM Project, we offered to provide some guidance to the HKU Team on the “PETase” bio brick as well as share (on their request) the PETase plasmid from last year’s 2020 CityU iGEM team. After plaiting it on an LB/amp plate, we selected a viable colony, cultured it in LB broth (10 mL), extracted the plasmid and then gave it to the HKU iGEM Team. We also provided the HKU team with the plasmid map and a snap gene file of the PETase plasmid. The HKU iGEM team thanked us profusely because it helped save them a significant amount of time and energy in constructing the PETase plasmid.

Figure 5. Handing over the PETase plasmid to the HKU Team (on 22nd July)

UCCKE Biology Research Team

Mentoring and Experimental Assistance to the UCCKE Biology Research Team

The United Christian College Kowloon East Biology Research Team contacted our Principal Team Advisor – Prof. Richard Kong – for his expertise in DNA Diagnostic Tools to seek his assistance and guidance with their iGEM project proposal. The UCCKE iGEM Team was aiming to develop a “Bacterial-based Detection Toolkit with Coupled Cell Proliferation Control” for biologists, where the detection of the target substance will restore normal cell proliferation in a genetically-modified strain of E. coli where several growth-related genes have been knocked out.

Figure 6. Prof. Kong in a scientific meeting with the UCCKE iGEM Team at CityU (on 8th Sept)

The CityU iGEM team held a face-to-face meeting with the UCCKE high-school team at the CityU Campus on 8th September 2021 to better understand the goals of their team project, the technical problems they encountered and offered suggestions for trouble shooting.

During the discussion with the UCCKE Team, Prof. Kong advised the team to consider the niche of the application of their project. He referred the UCCKE Team to the 2014 CityU iGEM Project – “FitColi: construction of a genetically-modified probiotic E. coli strain to enhance the uptake and conversion of excess fatty acids into DHA and EPA”. He commented that the UCCKE team’s aim to integrate a “cell growth control component” to their gene circuit would be useful in enhancing the growth of their E. coli, and the genes would be expressed only upon detecting the specific effector molecule (i.e., caffeine). Additionally, because the UCCKE team’s growth assay results showed a broad error bar, Prof. Kong advised their team to repeat the growth experiments to verify the reproducibility of their data. Our iGEM team also suggested that they conduct additional experiments on “leakage” expression and dose response assays at different concentrations of caffeine to further verify their data.

Figure 7. CityU-UCCKE Joint Team Meeting at the CityU campus (on 8th Sept)

Lastly, the UCCKE team shared some of the technical problems they were experiencing with plasmid transformation into E. coli cells. Although transformation of the plasmid (which encode for an MRP protein and Ampicllin resistance) into E. coli produced ampicillin-resistant colonies, colony PCR did not produced any MRP amplicon. However, PCR of the positive control (original plasmid) produced an MRP amplicon of the expected size. Moreover, checking for gene expression by induction of the Lac Operon on the plasmid with IPTG (which would produce a red fluorescence band if plasmid successfully transformed) turned out negative.

Our CityU team helped the UCCKE team repeat their plasmid transformation protocol in our lab at CityU, whereby the pUC19 plasmid was transformed into E. coli BL21 host cells followed by PCR amplification of the MRP gene using the TaKaRa Ex Taq polymerase. We successfully amplified the MRP gene as confirmed by gel electrophoresis. We further confirmed induction of the Lac operon using IPTG and detected the red fluorescence. The recombinant E. coli clone was then handed over to the UCCKE Team. In September, our CityU team invited the UCCKE Team and all other teams to participate in the “Synthetic Biology in the Modern World” International Talk Series organized by the CityU iGEM team.

Figure 8. Experiment on IPTG-induced MRP protein expression by a recombinant E. coli clone.

Figure 9. Handing over the recombiant MRP plasmid to the UCCKE representative (on 16th Sept)

The Hong Kong International School (HKIS) iGEM Team

Meeting with the HKIS iGEM Team

We held our first meeting with the Hong Kong International School iGEM Team on the 30th of July 2021. The meeting was a get to know you event where we met up with some impressive high school students taking part in the iGEM competition this year. We talked about each other’s projects, and we realized that the HKIS team, much like the HKUST team, were also tackling a very local problem with ambitious goals. They are aiming to develop a cheaper and faster detection and purification system for vibrio bacteria (as well as other pathogens) on oysters.

Figure 10. CityU-HKIS Zoom Meeting (on 30th July)

After an hour-long conversation, we wished each other good luck and decided to spread each of our project opinion surveys in our high school and university respectively for broader outreach. Furthermore, we had the pleasure to meet and talk with them at the HK iGEM Symposium, and we included them in our vlog of the 1-day Symposium.

The Siberian iGEM Team

Meeting with the Siberia iGEM Team – Leap2Brain

>We met with the Siberian iGEM Team – Leap2Brain - on the 3rd of August 2021 to widen our horizons and learn more about iGEM teams outside of Hong Kong. We found the “Leap2Brain” project to be one of the most ambitious iGEM projects for this year and we managed to form a good and friendly relationship with the Siberian team of the Novosibirsk State University.

>In our meeting we decided to collaborate in multiple ways – we prepared a Russian translation script of our “project opinion survey” and shared it with the Siberia team, and they in turn shared with us their “opinion survey” translated into English so that we could publicize it in Hong Kong on their behalf to help them get more responses. Additionally, they gave us their honest feedback and useful suggestions on our project presentation. For example, they pointed out to us that we have not provided the extraction protocol for our target variecolin metabolites from our fungal host given the fact that metabolite extraction was one of the key parts of our project since we are aiming to identify some potentially new medicinal compounds using a synthetic biology approach. On the other hand, the Siberian iGEM team were experiencing some technical issues with PCR and Western Blotting for protein visualization in their project. We shared with them a western blotting protocol that works well in our lab and gave them some tips to increase their success rate. We also shared with them information on our PCR protocol and the brand of Taq polymerase that worked well for us.

Figure 11. CityU-Siberia iGEM teams on a Zoom Meeting (on 3rd Aug)