Circular RNAs (circRNAs), formed by non-sequential back-splicing of pre-mRNA transcripts, are a wide- spread form of non-coding RNA in animal cells. Various function of natural existed circRNA revealed recently shows that circRNA can be used as powerful tools in future research and health care. However, there is no toolbox to generate circRNA up to date, which restrict the research and application of circRNA. Our project focus on the devices to cyclize specific part of RNA, aiming to start a circRNA revolution.
We designed three types of devices to cyclize the RNA based on the back-splicing mechanism:the "Ouroboros"(cyclizing device based on the inverted repeat sequence in the exon-flanking region), the "Cyclizers"(proteins that accelerate RNA cyclization, including "CyCli2er" and "LyCli2er")and the acRNA(ssRNA that accelerate RNA cyclization).
We experimentally validate that our device can accelerate the formation of cirRNA. We also measured the half-life time of the circRNA, which proved its stability. To solidity our experiment procedure, we designed a device to report the level of mir-21 concentration and modeled the steady state of proteins to confirm our design.
Apart from our lab work, we also took active part in the educational outreach activities which has had huge influences on and off campus. As sociable participants in iGEM, we have collaboration with NYU_Shanghai, ZJU-China and NCTU Formosa, and mentored a high school team WLSA_Fudan-Shanghai.
Regulating oncomiRs is essential to fight cancer! However, the ordinary RNA sponge can be easily degraded. So we give the sponge a shield: we cyclize it!
iGEM competition is based on standard parts, and we have modified most of our sequence to meet the RFC.
All of our parts to build the “Ouroboros” are all proved to be working as expected, as the testing experiment. Our mir-21 reporter also work as expected, which prove our design of mir-21 binding sites.
We also improved several parts, we develop several linker based on former linker. Apart from that, we use beta-globin intron to provide SA and SD sequence in our circular RNA device, which is inspired by the beta-globin intron parts.
We are working really hard to the outreach activities!
One of the major work we focus on is the popularization of science. Inspired by one of the major incident in the debating of transgenic technology in China, our team hold six successful lecture in our university to share the basic knowledge of different areas of biology. We are not satisfied with our achievements on the campus, so we search places outside our university to make more difference. We work hard to get the chance to give a speech to all the students in a high school; we explain the simple principle of transgenic food to the passengers and hand out leaflets on the street; we introduced our project to visitors in Shanghai Science&Technology Museum.
Besides all these activity, we also run the daily activity of Betalanffy Association, which is a famous community belonging to our iGEM team.
2015 Fudan is a fabulous team. We functioned specifically and managed every aspect of the iGEM competition within our team: project design, lab work, trouble-shooting, modeling, fundraising, human practice and wiki construction. With the generous help of our advisors and instructors, we brought here our final results. Also we offered our sincere thanks and acknowledgements for all the supporters involved in helping making a fabulous team. We do appreciate it.
We have shown our design details of three kinds of cyclizing device in the DESIGN page:the Ouroboros（cyclizing device based on the inverted repeat sequence in the exon-flanking region), the Cyclizer（proteins that accelerate RNA cyclization)and the acRNA(ssRNA that accelerate RNA cyclization).
All the three types of cyclizing devices we designed were actualized into sequence and parts, and we are currently working on testing and improve these devices.
We experimentally validate that our device can accelerate the formation of cirRNA.
We measured the half-life time of the circRNA produced by our device, which shows circRNA has significant higher stability.
We designed a device to report the level of mir-21 concentration to support our experiment.
We modeled the structure of proteins to confirm our “Cyclizer” design.
We test cyclization based on acRNA and find circRNA never documented before! We are now working on new experiments to investigating it!
Our devices provide huge possibility for cirRNA research and regulating oncomiRs, and we are still working to improve our device and develop our toolbox!
