A conditional knockout rat resource of mitochondrial protein-coding genes via a DdCBE-induced premature stop codon-Reference-Cited by-同舟云学术

A conditional knockout rat resource of mitochondrial protein-coding genes via a DdCBE-induced premature stop codon

Published:2023-04-14 Issue:15 Volume:9 Page:
ISSN:2375-2548
Container-title:Science Advances
language:en
Short-container-title:Sci. Adv.

Author:

Tan Lei¹^ORCID,Qi Xiaolong²^ORCID,Kong Weining³^ORCID,Jin Jiachuan⁴^ORCID,Lu Dan²^ORCID,Zhang Xu³^ORCID,Wang Yue¹,Wang Siting¹,Dong Wei³,Shi Xudong³,Chen Wei²,Wang Jianying¹,Li Keru²,Xie Yuan⁵,Gao Lijuan²,Guan Feifei³,Gao Kai³,Li Chaojun¹^ORCID,Wang Cheng¹⁵^ORCID,Hu Zhibin¹⁶⁷^ORCID,Zhang Lianfeng²⁸^ORCID,Guo Xuejiang¹^ORCID,Shen Bin¹⁶⁷⁹^ORCID,Ma Yuanwu²⁸¹⁰^ORCID

Affiliation:

1. State Key Laboratory of Reproductive Medicine, Women’s Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing Medical University, Nanjing, Jiangsu, China.

2. Key Laboratory of Human Disease Comparative Medicine, National Health Commission of China (NHC), Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences, Peking Union Medicine College, Beijing, China.

3. Beijing Engineering Research Center for Experimental Animal Models of Human Critical Diseases, Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences, Peking Union Medicine College, Beijing, China.

4. Center for Reproductive Medicine, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, China.

5. Department of Bioinformatics, School of Biomedical Engineering and Informatics, Nanjing Medical University, Nanjing, Jiangsu, China.

6. Center for Global Health, School of Public Health, Nanjing Medical University, Nanjing, Jiangsu, China.

7. Gusu School, Nanjing Medical University, Nanjing, Jiangsu, China.

8. Neuroscience center, Chinese Academy of Medical Sciences, Beijing, China.

9. Zhejiang Laboratory, Hangzhou, Zhejiang, China.

10. National Human Diseases Animal Model Resource Center, Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences, Peking Union Medicine College, Beijing, China.

Abstract

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.

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

Link

https://www.science.org/doi/pdf/10.1126/sciadv.adf2695

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