PUBLICATION

A knock-in strategy for editing human and zebrafish mitochondrial DNA using mito-CRISPR/Cas9 system

Authors
Bian, W.P., Chen, Y., Luo, J., Wang, C., Xie, S., Pei, D.
ID
ZDB-PUB-190409-1
Date
2019
Source
ACS synthetic biology   8(4): 621-632 (Journal)
Registered Authors
Bian, Wanping, Chen, Yanling, Luo, Juanjuan, Pei, Desheng, Wang, Chao, Xie, Shaolin
Keywords
none
MeSH Terms
  • Animals
  • CRISPR-Cas Systems/genetics*
  • Cell Line
  • DNA Repair/genetics
  • DNA, Mitochondrial/genetics*
  • DNA, Single-Stranded/genetics
  • Endonucleases/genetics
  • Gene Editing/methods
  • Gene Targeting/methods
  • HEK293 Cells
  • Homologous Recombination/genetics
  • Humans
  • Mitochondria/genetics
  • Mutagenesis/genetics
  • Transcription Activator-Like Effector Nucleases/genetics
  • Zebrafish/genetics*
  • Zinc Finger Nucleases/genetics
PubMed
30955321 Full text @ ACS Synth Biol
Abstract
The mitochondria DNA (mtDNA) editing tool, Zinc finger nucleases (ZFNs), Transcription activator-like effector nuclease (TALENs), and Clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9) system, is a promising approach for the treatment of mtDNA diseases by eliminating mutant mitochondrial genomes. However, there have been no reports of repairing the mutant mtDNA to date. Here, we first show a mito-CRISPR/Cas9 system that mito-Cas9 protein can specifically target mtDNA and reduce mtDNA copy number in both human cell and zebrafish. An exogenous single-stranded DNA (ssDNA) with short homologous arm can be accurately knocked into the targeting loci, and this mutagenesis could be steadily transmitted to F1 generation of zebrafish. Moreover, we found some major factors of nuclear DNA repairing could be significantly up-regulated by mito-CRISPR/Cas9 system. Taken together, our data suggested that the mito-CRISPR/Cas9 system could be a useful method to edit mtDNA, providing a potential therapy for the treatment of inherited mitochondrial diseases.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping