CRISPR/Cas9-mediated Mutagenesis at Microhomologous Regions of Human Mitochondrial Genome

Overview

Journal Sci China Life Sci

Specialties Biology
Science

Date 2021 Jan 9

PMID 33420919

Citations 12

Authors

Bang Wang

Xiujuan Lv

Yufei Wang

Zhibo Wang

Qi Liu

Bin Lu

Yong Liu

Feng Gu

Affiliations

Soon will be listed here.

Abstract

Genetic manipulation of mitochondrial DNA (mtDNA) could be harnessed for deciphering the gene function of mitochondria; it also acts as a promising approach for the therapeutic correction of pathogenic mutation in mtDNA. However, there is still a lack of direct evidence showing the edited mutagenesis within human mtDNA by clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9). Here, using engineered CRISPR/Cas9, we observed numerous insertion/deletion (InDel) events at several mtDNA microhomologous regions, which were triggered specifically by double-strand break (DSB) lesions within mtDNA. InDel mutagenesis was significantly improved by sgRNA multiplexing and a DSB repair inhibitor, iniparib, demonstrating the evidence of rewiring DSB repair status to manipulate mtDNA using CRISPR/Cas9. These findings would provide novel insights into mtDNA mutagenesis and mitochondrial gene therapy for diseases involving pathogenic mtDNA.

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