학술논문
Precise in vivo genome editing via single homology arm donor mediated intron-targeting gene integration for genetic disease correction
Document Type
article
Author
Suzuki, Keiichiro; Yamamoto, Mako; Hernandez-Benitez, Reyna; Li, Zhe; Wei, Christopher; Soligalla, Rupa Devi; Aizawa, Emi; Hatanaka, Fumiyuki; Kurita, Masakazu; Reddy, Pradeep; Ocampo, Alejandro; Hishida, Tomoaki; Sakurai, Masahiro; Nemeth, Amy N; Nuñez Delicado, Estrella; Campistol, Josep M; Magistretti, Pierre; Guillen, Pedro; Rodriguez Esteban, Concepcion; Gong, Jianhui; Yuan, Yilin; Gu, Ying; Liu, Guang-Hui; López-Otín, Carlos; Wu, Jun; Zhang, Kun; Izpisua Belmonte, Juan Carlos
Source
Cell Research. 29(10)
Subject
Language
Abstract
In vivo genome editing represents a powerful strategy for both understanding basic biology and treating inherited diseases. However, it remains a challenge to develop universal and efficient in vivo genome-editing tools for tissues that comprise diverse cell types in either a dividing or non-dividing state. Here, we describe a versatile in vivo gene knock-in methodology that enables the targeting of a broad range of mutations and cell types through the insertion of a minigene at an intron of the target gene locus using an intracellularly linearized single homology arm donor. As a proof-of-concept, we focused on a mouse model of premature-aging caused by a dominant point mutation, which is difficult to repair using existing in vivo genome-editing tools. Systemic treatment using our new method ameliorated aging-associated phenotypes and extended animal lifespan, thus highlighting the potential of this methodology for a broad range of in vivo genome-editing applications.