Team:BNU-China/Attributions/TeamAttributions
Accomplishment
With instruction of human practices, BNU-China set a goal of fixing the obesity epidemic by developing a synthetic leanness-promoting microbe colonized in human intestine. We have constructed and verified seven components respectively in wet lab, then integrated them into one system to demonstrate functions of multi-component engineered microbe. At the same time, we kept refining our project by constant human practice and modelling.
Training
After recruitment, training, and tested by team members of BNU-China 2018, 22 undergraduates stood out from 86 applicants. The training lasted from 12th to 18th of November, 2018, during which we learnt theoretical knowledge, analyzed excellent iGEM projects, brainstormed and gave presentation. With offered study materials, we taught ourselves in winter vacation, submitting study notes at the beginning of the next semester.
Brainstorming
After establishment of BNU-China 2019, team members of BNU-China began brainstorming on March 31st and primarily determined the project on June 5th. Thereafter we have been constantly completing our project until June 27th, before getting down to wet lab.
Wetlab
We started our wet lab experiment on June 28th 2019. Under the arrangement of team leaders, 14 wet lab team members were divided into several groups to construct different components. With the development of our project, we regrouped once for further research.
Acetate production
Zining Cao, student of BNU, proposed the idea of overproducing acetate, found the key enzymes, phosphotransacetylase (PTA) and acetokinase (ACK) in acetate production.
Yiyun Zhang, was responsible for finding foundational literatures that led to our innovation of acetate production.
Zhe Feng, Yiyun Zhang, student leaders of BNU-China, constructed a pathway that enhances acetate production and verified the feasibility. Furthermore, they combined acetate production and β-oxidation pathways and transferred them into a same strain to prove that the yield of acetate can be boosted by β-oxidation of fatty acids.
β-oxidation
Banglong Wang, Miao Liu, Xiaojing Li, students of BNU, designed and constructed the β-oxidation of fatty acids pathway, verified the functions of acyl-CoA synthetase (FadD) and acyl-CoA dehydrogenase (FadE) respectively, and helped measure the consumption of fatty acids after coexpressing acetate production and β-oxidation pathways. Banglong Wang was in charge of this group and designed the plasmids. Xiaojing Li found the key enzymes.
Yiyun Zhang, found the literatures that reveals the effect of high fat diet on obesity, supporting the idea of having our engineered bacteria consume the extra in-taken fat.
Bilateral switch
Zhe Feng, who designed the methodology, helped in the initial plasmids design, optimized experiments, purposed usage of degradation tag and the appropriate expression proportion between integrase (Bxb1 gp35) and excisionase (Bxb1 gp47).
Zhuoyan Hai, Qi Wang, Dake Gao, Banglong Wang, Miao Liu, Xiaojing Li, students of BNU, constructed the recombinase system, and tested the efficiency of gp35 and gp35&gp47 co-expression systems respectively. Zhuoyan Hai and Banglong Wang were in charge of two parallel groups and designed the plasmids.
Di Huang, Dake Gao, students of BNU, verified the effect of degradation tag RepA, enlightening a way to boost the efficiency of bilateral switch.
Di Huang, Dake Gao, Chunxu Han, Qi Wang, students of BNU, were involved in confirming the validity of the recombinase system as a “bilateral” switch and whether our improved parts/methods work. Di Huang was in charge of this group and designed two expression vectors. Dake Gao designed two report plasmids.
Yiyun Zhang, Zhuoyan Hai found foundational literatures about recombinase system, clarifying the mechanisms.
Digestion sensors
Zining Cao, Peijie Zhang, Yuzhe Zang, students of BNU, found and verified five promoters sensitive to digestion signals, including glucose inhibitory promoter rpoH, deoxycholate activated promoter micF, glycocholate activated promoter osmY, acyl-CoA activated promoter fadD, and acyl-CoA inhibitory promoter fabB.
Colonization
Zhe Feng, proposed several ways to colonize engineered bacteria inside human intestine.
Safety
Xiao Chen, Chunxu Han, Yuzhe Zang, Yiyun Zhang, Dake Gao, students of BNU, took part in the verification of induced suicide pathway and kill switch. Xiao Chen took responsibility of this team, found applicative enzymes and designed the plasmids.
Zhe Feng helped design verification experiments.
Integration
Zhe Feng, Yiyun Zhang, integrated all components into one plasmid and verified its functions. Zhe Feng designed the plasmid and all experimental methods.
Modelling
Tianxin Lv, student of BNU, constructed glucose concentration model and recombinase model, one helped to validate whether the sensitivity of rpoH works as expected, the other enlightened two modifications to increase the efficiency of bilateral switch.
Xinglei Ma, student of BNU, constructed the β-oxidation model, proving overexpressing fadD works more efficiently than fadE.
Yiyun Zhang, constructed protein docking model of gp35 and gp47, to identify the most suitable loci for adding degradation tag.
Yiyun Zhang, Zhe Feng, helped find biological parameters.
Art
Xiaodi Chen, Jiayin Wang, Die Hu, students of BNU, worked on the art of the project, including posters, logos, design of team uniform, graphics and beautification of wiki.
Wiki
Yajing Wang, Qingyun Xiong, Han Zhang, students of BNU, designed and completed the wiki structure, uploaded contents to our wiki.
Han Zhang, was in charge of wiki gruop, effectively managed work division and cooperation, helped to provide suggestions and ideas for art elements design.
Zhe Feng, Yiyun Zhang, took responsibility for writing articles presented in Wiki.
Collaborations
Zhe Feng was in charge of connecting other teams, organizing meetup and collaborations. Yiyun Zhang, Zining Cao, Peijie Zhang participated, too.
Human Practices
Xiaojing Li, sent questionnaire about high fat diet.
Zhuoyan Hai, sent questionnaire about probiotic products.
Zining Cao, Peijie Zhang, Yuzhe Zang, did market research on probiotics.
Xiaojing Li, Zhe Feng, Yiyun Zhang, Banglong Wang, Miao Liu, interviewed the sanitary bureau director of Dongcheng District in Beijing.
Zhe Feng, Yiyun Zhang, Zining Cao, Peijie Zhang, Yuzhe Zang, interviewed Prof. Yinyu Sun and Prof. Youhe Gao.
Zhe Feng, Zining Cao, Peijie Zhang, Yuzhe Zang, interviewed Prof. Yonggong Zhai.
Organization
• Treasury
Zhe Feng, Yiyun Zhang, were responsible for all financial management and controlled the budgets and invoices received.
• Coordination
Zhe Feng, Yiyun Zhang, as team leaders, scheduled experiments, organized and coordinated the whole team throughout the project.
Communication
All team members participated in hosting and attended Beijing regional meetup at Beijing Normal University. Di Huang and Zhuoyan Hai gave a presentation.
Zhe Feng, Yiyun Zhang, Zining Cao, Peijie Zhang, Di Huang, Zhuoyan Hai, attended the meetup at Beijing Institute of Technology. Zining Cao and Peijie Zhang gave a presentation.
Zhe Feng, Yiyun Zhang, Zining Cao, Banglong Wang, Zhuoyan Hai, attended the sixth Conference of China iGEMer Community in Shenzhen. Yiyun Zhang and Banglong Wang gave a presentation.
Project Support and Advice
Jinbo Chen, primary PI, was in charge of fundraising and the organization of BNU-China, offered us advices and financial support.
Hao Xiaoran, secondary PI, gave lab safety training, supported the team in experiments and devices, managed our laboratory space and taught us techniques we applied in the development of the biological part.
Prof. Xudong Zhu, Prof. Dong Yang, instructors of BNU-China, offered inspirations and guidance, gave us general advices in all the aspects of the whole project, and helped optimize experiments and texts.
Prof. Sen Li, Prof. Benqiong Xiang, instructors of BNU-China, provided financial support and coordinated the financial bureaucracy at the university.
Advisor
Yang Miao, advisor of BNU-China, gave us experimental training, made suggestions of the whole project, provided guidance of coordinating the financial bureaucracy, and booked hotel and air tickets.