Mitochondrial DNA editing system based on TALE assembly

Abstract

The invention belongs to the technical field of gene editing, and discloses a mitochondrial DNA editing system based on TALE assembly. The editing system comprises an RVD library, a mitochondrial localization skeleton carrier and a cell nucleus localization skeleton carrier, and the RVD library comprises at least 192 RVD modules including 160 double RVD modules, 16 single RVD modules and 16 half RVD modules. According to the editing system, the assembly efficiency of a TALE module can be remarkably improved, and the assembly cost and time of the TALE module can be remarkably reduced. According to the mitochondrial DNA editing system based on the TALE assembly, the assembly method is easy to operate and suitable for application and popularization. By means of the method, mitochondrial DNA editing can be rapidly and efficiently achieved, and the method is used for manufacturing various animal models so as to simulate diseases caused by human mitochondrial DNA mutation; the method can be potentially applied to gene therapy of mitochondrial DNA mutation diseases.

Description

technical field [0001] The invention belongs to the technical field of gene editing, in particular to a mitochondrial DNA editing system based on TALE assembly. Background technique [0002] Transcription activator-like effector (TALE) is the core module of second-generation gene editing tools, which is mainly composed of several 34 amino acid repeating units, and uses two repeating variable residues (RVD, repeat-variable di-residue) specifically recognizes DNA sequences. A variety of methods have been reported for the assembly of TALE modules, but most of these methods are tedious, time-consuming, laborious, and expensive, so how to efficiently assemble customized TALEs has become a difficulty in its promotion and application. [0003] Mitochondrial diseases are a class of major diseases characterized by abnormal mitochondrial function. Mitochondrial diseases are often caused by nuclear gene mutations and mitochondrial DNA mutations, and mitochondrial DNA mutations usuall...

AI Technical Summary

Problems solved by technology

Using ZFN and TALEN plus mitochondrial localization signal (MTS), the mutant mitochondrial DNA can be cut, causing the mitochondrial DNA with double-strand breaks to be degraded, but this method cannot be used to construct mitochondrial DNA mutant animal models, and use For the repair of mitochondrial DNA mutations, it has great limitations

CRISPR / Cas9 cannot be used for mitochondrial DNA editing due to the inability of sgRNAs to enter mitochondria

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