A product and fusion protein for treating and/or preventing beta hemoglobinopathies

Abstract

The application discloses a product and fusion protein for treating and / or preventing beta hemoglobinopathies. The fusion protein includes the DNA binding domain of Rad51, a mutant of cytosine deaminase APOBEC3A and a nuclease; Amide mutation to glycine. The product includes the coding gene of the fusion protein and the delivery vector of sgRNA, and the sgRNA guides the fusion protein to perform single-base gene editing on the promoter regions of HBG1 and HBG2 in target cells. This application can greatly improve the efficiency of single-base gene editing, more precisely and efficiently target HBG1 and HBG2 promoter regions -117 to generate mutations from G to A, thereby activating the expression of γ-globin, and providing clinical treatment for β-hemoglobin The disease provides a more precise and efficient treatment strategy.

Description

technical field

[0001] The invention relates to the technical field of genetic engineering, in particular to a product and fusion protein for treating and / or preventing beta hemoglobinopathies. 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 fro...

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