Quantitative computational analysis-based site-specific integrases in precision gene editing

Site-specific integrases have shown great potential for application in the field of gene editing, making them promising to assist in the efficient and precise insertion of DNA fragments.In this paper, site-specific recombinases are used as an entry point to mediate recombination between sites using site-specific integrases.The formation and characteristics of the tyrosine recombinase family and the serine recombinase family are analyzed separately.Subsequently, the different structures and characteristics of the recognition sites on catalytic reactions of the two types of recombinase families are highlighted.Under this theoretical framework, pNZTS01 temperature-sensitive plasmid, pNZTS-PnisA-dCas9-pmCDA1-DBtsI empty plasmid and pNZTS-CRISPR-cDBE-DBtsI empty plasmid were constructed to establish the temperature-sensitive backbone plasmid.Meanwhile, suitable targets were selected to synthesize sgRNA expression cassettes and establish base editor plasmids, thus proposing a gene editing method based on site-specific recombinase.In the in vivo therapeutic effect observation experiment of RNA nanococoon in mice, RNCOs-D induced the highest tumor growth inhibition rate (~78%), which verified that the gene editing based on site-specific recombinase could effectively inhibit tumor growth.Index Terms site-specific integrase, gene editing, base editor plasmid,