Fusion protein for improving gene editing efficiency and application thereof

Abstract

The invention discloses a fusion protein for improving gene editing efficiency and application thereof. The fusion protein comprises a single-stranded DNA binding protein functional domain, nucleoside deaminase and nuclease. According to CBEs, in the process of carrying out C-G to T-A base conversion, nucleoside deaminase such as cytosine deaminase takes single-stranded DNA as a substrate to carry out deamination, and a single-stranded DNA binding protein functional domain is fused on fusion protein of nucleoside deaminase and nuclease, so that the opportunity that the single-stranded DNA is exposed to nucleoside deaminase is greatly increased, and the base editing efficiency is obviously improved. The single-base gene editing technology is improved in a breakthrough manner, and the application of the fusion protein in the aspects of gene editing, gene therapy, cell therapy, animal model manufacturing, crop genetic breeding and the like can be greatly promoted.

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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