High-safety base editor and construction method and application thereof

• Main Authors: ZHANG SHUQIAN, CHENG TIANLIN
• Format: Patent
• Language: Chinese; English
• Published: 02.02.2024
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention relates to a high-safety base editor as well as a construction method and application thereof. The base editor is a new fusion protein obtained by fusing a cytidine deaminase mutant at different sites of mutant nuclease. Wherein the cytidine deaminase mutant is a new protein obtained by mutating the 173rd amino acid of YE1 from leucine to glutamine, the YE1 is a mutant obtained by mutating the 90th amino acid of APOBEC1 from tryptophan to tyrosine and mutating the 126 amino acid of APOBEC1 from arginine to glutamic acid, and the sequence of the APOBEC1 is as shown in SEQ.ID NO. 1. A novel cytidine deaminase mutant is developed and screened, the safety of a CBE tool constructed on the basis of the novel cytidine deaminase mutant is obviously improved, and the influence on gene expression, RNA splicing, DNA damage and mitochondrial functions is obviously reduced. According to the method, the system optimization of the safety of the single-base editing tool is realized. 本发明涉及一种安全性高的碱基编辑器及其构建方法与应用。碱