A fusion protein for improving gene editing efficiency and its application

Abstract

The present application discloses a fusion protein for improving gene editing efficiency and its application. The fusion protein includes single-stranded DNA binding protein functional domain, nucleoside deaminase and nuclease. According to the C-G to T-A base conversion process performed by CBEs, nucleoside deaminases such as cytosine deaminase deaminate single-stranded DNA as a substrate, by fusion of nucleoside deaminase and nuclease The single-stranded DNA binding protein functional domain is fused to the protein, which greatly increases the chance of single-stranded DNA being exposed to nucleoside deaminase, thereby significantly improving the efficiency of base editing. The present invention makes a breakthrough improvement on the single-base gene editing technology, which can greatly promote its application in gene editing, gene therapy, cell therapy, animal model making, crop genetics and breeding, etc.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a fusion protein for improving gene editing efficiency and its application. Background technique [0002] Since 2013, a new generation of gene editing technology represented by CRISPR / Cas9 has entered various experiments in the field of biology, changing the traditional means of gene manipulation. In April 2016, David Liu's laboratory first reported the single-base gene editing technology. After that, other types of single-base gene editing technology based on the principle of cytosine deaminase (such as cytosine deaminase from lamprey and human) Ammases are fused to dCas9 or Cas9n in different ways) have also been reported successively. It uses CRISPR / Cas9 derived from Streptococcus pyogenes spCas9 to use NGG as PAM and recognizes and specifically binds DNA to achieve a single base mutation from C to T or G to A upstream of NGG. [0003] Single-base gene editing technology has bee...

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Problems solved by technology

These methods all improve their efficiency to a certain extent, but are relatively limited.

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