DdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome

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 bas...

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Autores principales:	Jiayin Guo, Xiaoxu Chen, Zhiwei Liu, Haifeng Sun, Yu Zhou, Yichen Dai, Yu'e Ma, Lei He, Xuezhen Qian, Jianying Wang, Jie Zhang, Yichen Zhu, Jun Zhang, Bin Shen, Fei Zhou
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2022
Materias:	DdCBE mtDNA base editing mouse model mitochondrial disorder Therapeutics. Pharmacology RM1-950
Acceso en línea:	https://doaj.org/article/d0efd2d09726457fb6ad0eba937a0219
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spelling	oai:doaj.org-article:d0efd2d09726457fb6ad0eba937a02192021-12-04T04:33:42ZDdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome2162-253110.1016/j.omtn.2021.11.016https://doaj.org/article/d0efd2d09726457fb6ad0eba937a02192022-03-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2162253121002912https://doaj.org/toc/2162-2531Critical 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.Jiayin GuoXiaoxu ChenZhiwei LiuHaifeng SunYu ZhouYichen DaiYu'e MaLei HeXuezhen QianJianying WangJie ZhangYichen ZhuJun ZhangBin ShenFei ZhouElsevierarticleDdCBEmtDNAbase editingmouse modelmitochondrial disorderTherapeutics. PharmacologyRM1-950ENMolecular Therapy: Nucleic Acids, Vol 27, Iss , Pp 73-80 (2022)
institution	DOAJ
collection	DOAJ
language	EN
topic	DdCBE mtDNA base editing mouse model mitochondrial disorder Therapeutics. Pharmacology RM1-950
spellingShingle	DdCBE mtDNA base editing mouse model mitochondrial disorder Therapeutics. Pharmacology RM1-950 Jiayin Guo Xiaoxu Chen Zhiwei Liu Haifeng Sun Yu Zhou Yichen Dai Yu'e Ma Lei He Xuezhen Qian Jianying Wang Jie Zhang Yichen Zhu Jun Zhang Bin Shen Fei Zhou DdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome
description	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.
format	article
author	Jiayin Guo Xiaoxu Chen Zhiwei Liu Haifeng Sun Yu Zhou Yichen Dai Yu'e Ma Lei He Xuezhen Qian Jianying Wang Jie Zhang Yichen Zhu Jun Zhang Bin Shen Fei Zhou
author_facet	Jiayin Guo Xiaoxu Chen Zhiwei Liu Haifeng Sun Yu Zhou Yichen Dai Yu'e Ma Lei He Xuezhen Qian Jianying Wang Jie Zhang Yichen Zhu Jun Zhang Bin Shen Fei Zhou
author_sort	Jiayin Guo
title	DdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome
title_short	DdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome
title_full	DdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome
title_fullStr	DdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome
title_full_unstemmed	DdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome
title_sort	ddcbe mediates efficient and inheritable modifications in mouse mitochondrial genome
publisher	Elsevier
publishDate	2022
url	https://doaj.org/article/d0efd2d09726457fb6ad0eba937a0219
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_version_	1718372996488364032

DdCBE mediates efficient and inheritable modifications in mouse mitochondrial genome

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