Precise in vivo genome editing via single homology arm donor mediated intron-targeting gene integration for genetic disease correction.

Journal: 
Cell Res
Publication Year: 
2019
Authors: 
Keiichiro Suzuki
Mako Yamamoto
Reyna Hernandez-Benitez
Zhe Li
Christopher Wei
Rupa Devi Soligalla
Emi Aizawa
Fumiyuki Hatanaka
Masakazu Kurita
Pradeep Reddy
Alejandro Ocampo
Tomoaki Hishida
Masahiro Sakurai
Amy N Nemeth
Estrella Nunez Delicado
Josep M Campistol
Pierre Magistretti
Pedro Guillen
Concepcion Rodriguez Esteban
Jianhui Gong
Yilin Yuan
Ying Gu
Guang-Hui Liu
Carlos Lopez-Otin
Jun Wu
Kun Zhang
Juan Carlos Izpisua Belmonte
PubMed link: 
31444470
Public Summary: 
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.
Scientific 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.