The CRISPR/Cas9 Genome Editing System Application And Target Protein REV1 Mitochondrial Function Identification

Consideration of the importance of mitochondria which play significant roles in metabolism and cell differentiation, apoptosis, autophagy, and there exsit links between mitochondrial function disorders and the diseases which include cancers as well, the scientists have been paying much attention to mitochondrial DNA damage repair. The translesion DNA synthesis (TLS) is one of those mechanisms that refer to the nucleus DNA damage repair, it can bypass the stalled replication forks by replacing error-free polymerases with error-prone polymerases to prevent cells from dearth. Although there were researchers finding out that RAD 18 which was an important nuclear TLS protein could exist in mitochondrial components in vitro, there were no evidences that it might involve in mitochondrial TLS. Our lab is interested in REV1 which is a dCMP transferase and TLS Y-family polymerase and act as a scaffold to recruit other TLS proteins, so we did some researches to identify the functions that REV1 acted on mitochondria to explore whether TLS could exist in mitochondria as another mtDNA damage repair pathway or some views at least. It is very popular that genome-editing technologies studied in life science which commonly use zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR related protein 9 (Cas9) which act in bacterial adapted immune system. The application of the economical and effective CRISPR/Cas9 system can knock out the REV1 gene for a deeper research and provide experiences for our lab to reprogramed other target proteins.