|ZFIN ID: ZDB-PUB-190409-1|
A knock-in strategy for editing human and zebrafish mitochondrial DNA using mito-CRISPR/Cas9 system
Bian, W.P., Chen, Y., Luo, J., Wang, C., Xie, S., Pei, D.
|Source:||ACS synthetic biology 8(4): 621-632 (Journal)|
|Registered Authors:||Bian, Wanping, Chen, Yanling, Luo, Juanjuan, Pei, Desheng, Wang, Chao, Xie, Shaolin|
|PubMed:||30955321 Full text @ ACS Synth Biol|
Bian, W.P., Chen, Y., Luo, J., Wang, C., Xie, S., Pei, D. (2019) A knock-in strategy for editing human and zebrafish mitochondrial DNA using mito-CRISPR/Cas9 system. ACS synthetic biology. 8(4):621-632.
ABSTRACTThe 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.