Team:CPU CHINA/Collaborations
COLLABORATION
CONFERENCES AND MEETUPS
THE CONFERENCE OF NANJING iGEM TEAMS
On May 5th, 2021, the conference of Nanjing iGEM teams hosted by NJU-China was held in the Nanjing University. We were invited to participate in this very collaborative event in which we were able to acquire actual feedback of project design from others. At the meeting, each team made a brief introduction to their projects and future plans. Nanjing iGEM Association, a regional cooperation and exchange organization for iGEM teams in Nanjing, was officially established after the meeting.
Fig. 1 Members of CPU_CHINA presenting project background
Fig. 2 Group photo of CPU_CHINA participants during the conference of Nanjing iGEM teams
Fig. 3 Group photo of all the participants of the conference of Nanjing iGEM teams.
We gained suggestions that we shall further determine the authenticity of H2O2-producing hypothesis of laccase before we decided to implement it into experiments. In addition, we had received several new cooperation intentions with other iGEM teams. As a result, we planned to team up with NAU-China for future cooperations. This led to the launch of an educational event, i.e. "Into China, Into iGEM". We will put more explanation about this event later in this page.
YEAST MEETUP
As we selected Pichia pastoris as our engineered chassis, it is of vital importance that we acquire further insights of how to utilize this organism correctly and properly. Therefore, on August 22nd, 2021, we participated in the virtual Yeast Meetup hosted by UESTC-China. We shared our experience and specific details about culturing and fermenting Pichia pastoris.
Fig. 4 Participants of the virtual Yeast Meetup.
As the host of this meetup, UESTC-China shared their encyclopedical knowledge about summarized tips and cautions for using P.pastoris as the engineered chassis. In particular, their share of detecting miscellaneous bacterial infection in cultures of P.pastoris with centrifugation reminded us of an abnormal phenomenon we discovered when preparing P.pastoris competent cell, as the contaminated P.pastoris culture was not able to condense tightly at the bottom of centrifuge tube after centrifuging at 4000 rpm for 10 min. Using this abnormal competent cell for electrotransformation may lead to the failure of acquiring recombinant single cell colonies.
In this meetup, we found that a lot of teams were puzzled on how to perform yeast colony PCR with a higher success rate. They tried many kinds of yeast colony pretreatments such as adding lyticase or fast freezing-thawing, but with unstable success rate. Therefore, we provided our yeast colony PCR protocol of using 1 μl diluted colony sample (simply picks a portion of one single colony and dilutes the cells with 10μl ddH2O) as our template source, then setting a pre-denaturation stage at 98℃ for 10 min before switching to the normal colony PCR program.
Fig. 5 One of our Powerpoint slides showing protocols of plasmid linerization and pretreatment before electrotransformation.
As a result, we benefited a lot from discussing various problems we encountered when dealing with this organism. We also got the opportunity of sharing many unexpected "surprises" we experienced during culturing P. pastoris. In addition, we established a further cooperation intention with team UESTC-China.
THE 8th CCiC (CONFERENCE OF CHINA iGEMer COMMUNITY)
The Conference of China iGEMer Community (CCiC) is the biggest annual event for iGEMers from different teams around China to gather together, where participants are able to display their project, to exchange ideas and suggestions, as well as to look for of partners that share similar aspirations. In 2021, despite difficulties caused by the COVID-19 pandemic, we still made our way to the 8th CCiC hosted in Shanghai from August 27 to 29.
The conference had invited several honored guests ranging from prestigious academicians to successful entrepreneurs to deliver presentations regarding understanding of the developmental prospects of synthetic biology, personal experiences and field insights, and certain cautions of conducting an iGEM project. Also, during poster sessions, we made visits to other teams. Particularly, members from DUT_China welcomed us warmly. Similar in project goal, we had constructive conversations with each other and further determined our partner relationship.
Fig. 6 The photograph of team members from DUT_China and CPU_CHINA at CCiC8 venue.
We made a brief report of the design and progress of our project. Two judges informed us with vital feedbacks. What's more, during the period of online Q&A session, Huandi Xu, the president of CCiC executive committee, evaluated our project and provided us with helpful advice on improving presentation effectiveness. In respond to all these feedbacks, firstly, we need to better specify the application scenarios of our project, so that we can further modify our product with pertinence; secondly, we shall focus on proving or demonstrating that our protein-nucleic acid complex could converge the advantages of the three PE-degrading parts to exert a more significant functional improvement when compared with independently applying the parts into use.
At CCiC8, we stood out from the crowd of teams and entered the finalist of Best Presentation Awards.
Fig. 7 The "Best Presentation Finalist" medal
CRISPR/Cas COMMUNICATION MEETING
Since CRISPR-related techniques were included in our project, on September 12th, we were invited to attend the virtual CRISPR/Cas Communication Meeting hosted by team Tianjin.
Fig. 8 Ongoing scene of CRISPR/Cas communication meeting
During the meeting, we had an in-depth exchange with teams using the CRISPR/Cas9 system, discussed and resolved the problems that each team encountered. We received assistance and advice from team Tianjin & NEFU_China, who suggested us to perform SDS-PAGE of lower separation gel concentrations to characterize our SpyCatcher/SpyTag linkage system. The separation of our multiplex protein system was significantly improved after we changed the conditions. What's more, they also suggested that separations could be performed by adapting pull-down experiments using an altered label.
After the meeting, we worked together on the writing of
ROUND TABLE CLUB: PLASTICASE
In late spring and early summer, team CPU_CHINA, as the organizer, united many iGEM teams to establish Plasticase--a community dedicated in solving the problem of plastic pollution using approaches derived from synthetic biology. From May to September, members of Plasticase shared cutting-edge information, looked for potential partners, and put forward solutions regarding various problems.
Fig. 9 Teams consist of Plasticase Confederation
In order to have a deeper understanding between every team, we decided to launch a round table club which allowed teams to exchange projects and explore opportunities for in-depth cooperation.
Fig. 10 Participants of Round Table Club
With the help of IvyMaker-China, we invited Dr. Huiming Xiang from Shanghai Environment Group Company Limited to introduce the migration and hazards of microplastics in the environment to us. Later, iGEMers from BJEA_China, BUCT-China, CPU_CHINA, DUT_China, IvyMaker-China, KEYSTONE, SMS_Shenzhen and TJUSLS_China introduced their respective projects. As judges, Prof. Meiling Lu and Prof. Meirong Chen from China Pharmaceutical University also put forward their own suggestions and opinions. Whether it is a member of wet lab or human practices, everyone had learned a lot.
The round table club had brought every member of Plasticase closer with each other. We believe that the deeper sense of belonging to Plasticase in every member would facilitate further discussions regarding progresses in the field of synthetic biology and plastic degradation during and after the iGEM competition.
For further details of Plasticase Confederation, see Plasticase.
TEAM COLLABORATIONS
DUT_China
Team DUT_China shared similar goals of attenuating plastic pollution with us. They contacted us after watching the playback of our project introduction at the conference of Nanjing iGEM teams. They invited us to attend Laccase Meeting held by team TU_Kaiserslautern 2020 on May 18th, where we discussed the aforementioned H2O2-producing hypothesis of laccase with team members of TU_Kaiserslautern.
Fig. 11 Participants of Laccase Meeting.
After the Laccase Meeting, several online meetups between the two teams were held around June for introducing respective projects, further discussing and exchanging details. We met with each other at the 8th CCiC as described above.
Fig. 12 Regular inter-team meetups between DUT_China and CPU_CHINA
Apart from the communications and assistances described above, DUT_China also provided valuable opportunities for us to reach out to experts in industrial community. They as well obtained inspirations from our project designs.
NAU-CHINA
We met team NAU-CHINA at the conference of Nanjing iGEM teams in May. They not only put forward vital advice for our project during the conference, but also showed the strongest willingness to collaborate with us after we met with each other.
Soon, on June 15th, the first online meeting was hosted between Human Practice members of the two teams.
Fig. 13 The Human Practices members of the two teams discussing formats of education activities.
We discussed ways of carrying out our educational activities and realized that uniting teams from all over the nation to join us may be a good idea. Therefore, with the aim of maximizing our collaborative partners, we sent invitational emails to participating teams located in different provinces of China. Eventually, a total of 17 iGEM teams were recruited. Furthermore, we determined the format and workflow of this very activity, namely "ICII" (For further reading of ICII, please check the description below, as well as our Education page). We decided to demonstrate our achievements by displaying briefings on a map, as well as making vlog clips, so that they would be more intuitive and vivid for people to understand.
In the following periods, we allocated our respective responsibilities, finished the first promotion article, and prompting preaching activities hosted by participating teams around the nation. In addition, scriptwriting and video clipping of different materials from various teams were also important jobs that we accomplished.
NEFU_China
Team NEFU_China contacted us in August hoping to be more familiar with the role and function of CRISPR/dCas9 system in our project design. Therefore, they hosted a short meeting and shared the project design with each other. We also provided several experimental suggestions on how to verify the assembly of dCas9 with sgRNAs and dsDNA.
Fig. 14 Participants of the short meeting between CPU_CHINA and NEFU_CHINA.
Later, during the online Q&A session of CCiC, NEFU_CHINA consulted us about difficulties they met of assembling their system in vivo**. We thought that their system was too large and sophisticated to be assembled in bacteria, so we suggested that they should tackle this problem by trying to verify its assembly in vitro, as we did in our project.
We were glad to hear from members of NEFU_CHINA on September 5th. They informed us that they listened to our advice and had successfully obtained components of their system in vitro, and they would try to assemble it in vitro.
Tianjin
Team Tianjin reached out to us after we mentioned the usage of CRISPR/dCas9 techniques in our project during the presentation stage of CCiC. We were subsequently invited to participate in the aforementioned CRISPR/Cas meeting they hosted, where several teams applying this technique gathered together to exchange experiences and share experiments.
During the meeting, the two teams exchanged academic suggestions with each other, and tried to work out solutions on several problems we encountered in our experiments. We also discussed difficulties and solutions with other participating teams. After the conference, we accomplished the CRISPER/Cas handbook, hoping to give experienced references for future teams.
After the meeting, the two teams continued to collaborate with each other throughout the rest of iGEM 2021 season (For more information on how the two teams help with each other, see Partnership page).
Fig. 15 Members of the two teams discussing issues about experiment progress.
Tongji_China
Team Tongji_China is the first team that we made communication with. Early on March 28th, with the help of the instructors from both teams, we met online to discuss literally everything about iGEM, from familiarizing ourselves with the competition calendar to brainstorming with each other for threads of project design.
Fig. 16 The first ever meetup of team CPU_CHINA with Tongji_China
At the initial meeting, the two teams determined that we shall both work on environmental projects this year, thus potential collaborations regarding projects was possible.
Later in May, members of Tongji_China made a visit to the campus of CPU_CHINA. Together, the two teams made further discussions on updates of respective project design and human practices plans. We learned that Tongji_China dedicated to reduce stinks produced by rotten food in kitchen garbage dustbins, thereby improving air quality. After the meeting, we showed our guests around our laboratory and different places in the campus.
Fig.17 Members of the Tongji_China introducing project background to team CPU_CHINA.
Since the two teams shared a similar focus on environmental issues, as we were conducting survey in the waste transfer station of Jiangning High-tech Zone, we interviewed staffs about current treatments for garbage stinks, and discussed the possibility of applying engineered strains to solve this problem. After that, we provided the related feedback to Tongji_China.
UESTC-China
We met team UESTC-China through China iGEMer Online Meetup back in May. After discovering that both teams used Pichia pastoris strain GS115 to express target protein, we engaged in further communication and collaboration, such as discussing the problems encountered in our experiments and carried out educational work in Human Practices.
We both encountered many problems when using GS115 to express our protein, such as contamination issues caused by unknown species of miscellaneous bacteria, plasmid DNA cross-pollution in yeast electroporation issues, as well as numerous puzzling observations caused by the mysterious expression profile of Pichia pastoris. Our common daily complaints towards this very chassis had brought the two teams closer together, facilitating lots of conversations and helping hands, and eventually, partnership. (For more information on how the two teams help with each other, see Partnership page)
In addition to assistances of experiment-related issues, we also communicated with each other regarding the filling of Safety Form, as well as participating in the virtual Yeast Meetup described above.
SPECIAL ISSUE: INTO CHINA, INTO iGEM (ICII)
Fig. 18 The participating teams are located in different regions of mainland China.click here to map
"ICII" refers to "Into China, Into iGEM". This is an Education & Collaboration activity jointly sponsored by our team and NAU-China. 18 iGEM teams from all over China were invited to participate. As the 18 teams were located in different provinces of China showed above, by motivating paticipant teams to deliver preachings regarding synthetic biology and team projects at local middle schools or science and technology museums, a national publicity of synthetic biology was achieved. During the preaching sessions held by different teams in different locations, we encouraged the specific preaching team to display project information and posters of other cooperant teams to realize a better exhibition effect. In addition, the exhibiting process of every team was documented by video clips for the production of a final promotion vlog.
[1] Attachment: Posters made by CPU_CHINA and NAU-CHINA; The vlog made by CPU_CHINA
[2] For further reading of ICII workflows and the preaching accomplished by CPU_CHINA, see (Education website)