Team:ZJU-China/Parts

This year, we designed a set of parts. They consisted of different parts collection and play a role in the different section of our project. All the parts had been submitted to the iGEM Registry following the BioBricks assembly standard. Here is a brief description of some of our parts and we divide them into several individual sections for presentation. For more detailed information about our parts, we highly recommend you to visit the corresponding parts page on the iGEM BioBricks Registry or turn to the Design page of our wiki.

New Parts

Name Length(bp) Type Function and description designer
BBa_K3730000 104 promoter sPrcn+T7+RBS Chen Jiou, Yang Linhe
BBa_K3730001 35 DNA rcn operator Chen Jiou, Yang Linhe
BBa_K3730007 143 DNA 2216-TLR9i Chen Jiou, Hu Longshuang
BBa_K3730012 296 composite parts miRNA sponge whose mRNA will be able to temporarily combine the free miRNA. Wang Yongyin
BBa_K3730013 220 composite parts A modified 3' UTR with miRNA combining sequence. Wang Yongyin
BBa_K3730014 1222 composite parts E1A-3'TR Wang Yongyin
BBa_K3730017 263 promoter human tert promoter Huang Guanrui
BBa_K3730018 397 promoter survivin promoter Huang Guanrui
BBa_K3730023 63 shRNA primer pEGFP-C1 assembling promoter hTERT targeting shRNA forward primer-2 Huang Guanrui
BBa_K3730024 63 shRNA primer pEGFP-C1 assembling promoter hTERT targeting shRNA reverse primer-2 Huang Guanrui
BBa_K3730025 63 shRNA primer pEGFP-C1 assembling VEGFA targeting shRNA forward primer-2 Huang Guanrui
BBa_K3730026 63 shRNA primer pEGFP-C1 assembling VEGFA targeting shRNA reverse primer-2 Huang Guanrui

Improved rcn operator

We had made improvement on BBa_K540001. The traditional rcn operator is a cobalt response brick. We changed its constitutive sequence into T7 promoter and remain the cobalt controlled part. By increasing the expression of repressor, we are able to improve the degree of induction. You can find more information about our brick in the registry pages of BBa_K3730000 and BBa_K3730001.

Immunocammouflage

BBa_K3730007, BBa_K3730008. These parts are applied to prove the effect of TLR9i, which is used for temporarily immunocammouflage to avoid early clearance to the virus by our immune system. The insertion of the sequence into the virus genome will make a difference.

Tumor targeting mechanism

These parts aim to guarantee the safety of the oncolytic virus. BBa_K3730017 and BBa_K3730018 are the tumor specific expression promoter, which regulate a high expression in cancer cells while low in normal cells. BBa_K3730014 is a composite part which acts as a suicide trigger. It s the untranslated regions of E1A including microRNA target sites. This design allows free microRNA to combine the E1A gene, leading to the reduction of the cell viability.

miRNA sponge

BBa_K3730009, BBa_K3730010. We had designed a special DNA sequence which aimed to regulate the suicide mechanism. The mRNA produced by it will be able to combine the specific types of free miRNA in the tumor cells. This combination will not affect the suicide switch mechanism in normal cells, while in tumor cells, the miRNA concentration will be too low to trigger the mechanism. This design allows our oncolytic virulence part to work safely without reduction of efficiency.

Oncolytic virulence part

BBa_K3730023, BBa_K3730024, BBa_K3730025, BBa_K3730026. We had designed several shRNAs which target the hTERT and VEGFA gene for oncolytic virulence. We had proved their efficiency and the results are presented in Result page.

ZJU-China ZJU-China