DdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome

Molecular Therapy: Nucleic Acids

Fei Zhou¹  Xiaoxu Chen²  Lei He³  Haifeng Sun³  Yichen Zhu³  Jun Zhang³  Xuezhen Qian³  Yichen Dai⁴  Jiayin Guo⁴  Zhiwei Liu⁴  Yu Zhou⁴  Jie Zhang⁴  Bin Shen⁴  Jianying Wang⁴  Yu'e Ma⁴ 

[1] Corresponding author: Jun Zhang, 101 Longmian Avenue, Nanjing 211166, China.;Gusu School, Nanjing Medical University, Nanjing 211166, China;Cambridge-Suda Genomic Resource Center, Jiangsu Key Laboratory of Neuropsychiatric Diseases Research, Medical College of Soochow University, Suzhou 215123, China;State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing 211166, China;

关键词: DdCBE;    mtDNA;    base editing;    mouse model;    mitochondrial disorder;   

DOI  :  

来源: DOAJ

【 摘 要 】

Critical mutations of mitochondrial DNA (mtDNA) generally lead to maternally inheritable diseases that affect multiple organs and systems; however, it was difficult to alter mtDNA in mammalian cells to intervene in or cure mitochondrial disorders. Recently, the discovery of DddA-derived cytosine base editor (DdCBE) enabled the precise manipulation of mtDNA. To test its feasibility for in vivo use, we selected several sites in mouse mtDNA as DdCBE targets to resemble the human pathogenic mtDNA G-to-A mutations. The efficiency of DdCBE-mediated mtDNA editing was first screened in mouse Neuro-2A cells and DdCBE pairs with the best performance were chosen for in vivo targeting. Microinjection of the mRNAs of DdCBE halves in the mouse zygotes or 2-cell embryo successfully generated edited founder mice with a base conversion rate ranging from 2.48% to 28.51%. When backcrossed with wild-type male mice, female founders were able to transmit the mutations to their offspring with different mutation loads. Off-target analyses demonstrated a high fidelity for DdCBE-mediated base editing in mouse mtDNA both in vitro and in vivo. Our study demonstrated that the DdCBE is feasible for generation of mtDNA mutation models to facilitate disease study and for potential treatment of mitochondrial disorders.

【 授权许可】

Unknown