Progress of Site-specific Integration of Large Exogenous DNA Fragments Based on Non-viral Methods
Site-specific integration of large DNA fragments based on non-viral methods are widely used in gene therapy and cell therapy because of their low cost,large capacity,non-immunogenicity,and the ability to avoid genotoxicity and oncogene expression.According to the mechanism,non-viral methods can be classified into three types,DNA damage repair-,recombinase-and transposase-based site-specific integration technologies.All three non-viral DNA targeting methods can be combined with the clustered regularly interspaced short palindromic repeats(CRISPR)and CRISPR associated proteins 9(Cas9)technology to quickly and easily direct exogenous DNA to the target DNA region.In this study,the research progress of the three non-viral-based DNA targeting methods and their applications are summarized to provide new ideas for the development of more efficient cell therapy systems.