A conditional knockout rat resource of mitochondrial protein-coding genes via a DdCBE-induced premature stop codon.
Lei TanXiao-Long QiWeining KongJiachuan JinDan LuXu ZhangYue WangSiting WangWei DongXudong ShiWei ChenJianying WangKeru LiYuan XieLijuan GaoFeifei GuanKai GaoChao-Jun LiCheng WangZhi-Bin HuLianfeng ZhangXue-Jiang GuoBin ShenYuanwu MaPublished in: Science advances (2023)
Hundreds of pathogenic variants of mitochondrial DNA (mtDNA) have been reported to cause mitochondrial diseases, which still lack effective treatments. It is a huge challenge to install these mutations one by one. We repurposed the DddA-derived cytosine base editor to incorporate a premature stop codon in the mtProtein-coding genes to ablate mitochondrial proteins encoded in the mtDNA (mtProteins) instead of installing pathogenic variants and generated a library of both cell and rat resources with mtProtein depletion. In vitro, we depleted 12 of 13 mtProtein-coding genes with high efficiency and specificity, resulting in decreased mtProtein levels and impaired oxidative phosphorylation. Moreover, we generated six conditional knockout rat strains to ablate mtProteins using Cre/loxP system. Mitochondrially encoded ATP synthase membrane subunit 8 and NADH:ubiquinone oxidoreductase core subunit 1 were specifically depleted in heart cells or neurons, resulting in heart failure or abnormal brain development. Our work provides cell and rat resources for studying the function of mtProtein-coding genes and therapeutic strategies.
Keyphrases
- copy number
- mitochondrial dna
- oxidative stress
- genome wide
- heart failure
- induced apoptosis
- high efficiency
- bioinformatics analysis
- diabetic rats
- dna methylation
- genome wide identification
- single cell
- escherichia coli
- cell therapy
- genome wide analysis
- spinal cord
- spinal cord injury
- cell cycle arrest
- left ventricular
- small molecule
- cell death
- endothelial cells
- amino acid
- stem cells
- endoplasmic reticulum stress
- high glucose
- protein protein
- multiple sclerosis
- stress induced