Team:Nanjing NFLS/Parts

Team:Nanjing_NFLS 2017-igem.org

Parts.


Go to "Parts" and "Improve" from bottom navigation bar.

Part name Short Description Long Description
BBa_K2597000 HOsite-TMCP (HO recognizable site, Cas9) BBa_K2597000 consists of Homothallic switching endonuclease (HO) recognizable site, Mini-TK promoter, Cas9 and poly A, as we named to be HOsite-TMCP. This part codes for Cas9 gene and expresses Cas9 protein, aiming to cut the telomeres of cancer cells under the guidance of telomerase-targeting sgRNA. HO recognizable site could be recognized by HO enzyme to produce the four-base overhang (3’-TTGT-5’), which would be recognized by telomerase and lead to the synthesis of double-stranded telomeric repeat sequence.
BBa_K2597001 TsgRNA-dCas9-VP64 BBa_K259701 consists of TsgRNA designed to target telomeric repeat sequence and artificial transcription factor dCas9-VP64 to activate transcription of effector gene on the vector. We cloned the linear TsgRNA fragments target telomere into plasmid pcDNA-dCas9-VP64 to generate plasmid TsgRNA-dCas9-VP64 for convenience of using and improvement in transfection efficiency. When transfected in cells with high telomerase activity, dCas9-VP64 could bind the telomeric repeat sequence synthesized by telomerase thus activate the expression of effector gene Cas9, leading to its cutting on telomeres and the apoptosis of cancer cells.
BBa_K2597002 Homothallic switching endonuclease (C1-HO) BBa_K259702 is constructed to express HO enzyme that could cut a blunt-ended effector containing HO recognizable site (HOsite-TMCP) to produce a stick end of 3’-TTGT-5’. The stick end could be recognized by telomerase, leading to the double-stranded telomeric repeat sequence synthesis.