Screening and Analysis of Potential Genes for DNA Damage Repair and Apoptotic Signal in iPSCs Based on CRISPR-Cas9 System

Clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) 9 system is a site-specific gene editing technology developed in recent years, which can perform various genetic operations on specific sites of the genome, including gene targeting, site-specific gene insertion, gene repair, etc. The technology has shown a broad application prospect in basic scientific research and clinical disease treatment. CRISPR-Cas9 system has great potential in the treatment of genetic tumors. How to quickly repair DNA damage and improve the targeting efficiency is the key technical bottleneck that needs to be broken through in its clinical application. Based on this, we analyzed the gene dynamic change of CRISPR-Cas9 system in iPSCs from RNA expression data (SRP151274). By analyzing the dynamic change process, we further analyzed the mechanism of homologous recombination repair, aimed to explore new genes to improve the efficiency of knockin for iPSCs. As a result, we found that after induced pluripotent stem cells were transferred by electroporation, reducing cell death and interfering with DNA damage repair signal may be an important way to improve efficiency. DNA-binding proteins (GO: 0051101), the negative regulation of cell proliferation (GO: 0045596), DNA damage response (GO: 0030330), cell morphogenesis involved in differentiation (GO: 0000904), central nervous system neuron differentiation (GO: 0021953), negative regulation of cell proliferation (GO: 0008285) were the main biological process after electroporation. At last, seven potential genes, which might be a key intervention point that mediated the CRISPR-Cas9 system, were selected from DNA damage repair signal and the apoptotic signal.

Automated summary

CRISPR-Cas9 system is a gene editing technology that can perform various genetic operations on specific sites of the genome, showing broad application prospects. By analyzing the dynamic changes of CRISPR-Cas9 system in iPSCs, new genes can be explored to improve knockin efficiency; reducing cell death and interfering with DNA damage repair signal may be important ways to improve efficiency when treating iPSCs.