Construction method of disease animal model and fusion protein

Abstract

The invention discloses a construction method of a disease animal model and a fusion protein. The fusion protein comprises a DNA (Deoxyribose Nucleic Acid) binding structural domain of the Rad51, cytosine deaminase APOBEC3A and nuclease. The method comprises the following steps: introducing the fusion protein and sgRNA into animal cells, and carrying out gene editing on a target gene. The invention provides a novel platform for efficiently generating the disease animal model, and the manufacturing process of different animal models is greatly promoted.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for constructing a disease animal model and a fusion protein. 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 (Streptococcus pyogenes) spCas9 uses NGG as PAM (spacer precursor adjacent motif) and recognizes and specifically binds DNA upstream of NGG to achieve a single base from C to T or G ...

AI Technical Summary

Problems solved by technology

The new single-base gene editing technology has attracted much attention for producing disease animal models with 100% efficiency. However, the existing single-base gene editing technology is usually C3-C8, and targeting C near the PAM region is not very effective.

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