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

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 Nuñez Delicado, Josep M. Campistol, Pierre Magistretti, Pedro Guillen, Concepcion Rodriguez Esteban, Jianhui GongYilin Yuan, Ying Gu, Guang Hui Liu, Carlos López-Otín, Jun Wu, Kun Zhang, Juan Carlos Izpisua Belmonte

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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.

Original languageEnglish (US)
Pages (from-to)804-819
Number of pages16
JournalCell Research
Volume29
Issue number10
DOIs
StatePublished - Oct 1 2019

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ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Suzuki, K., Yamamoto, M., Hernandez-Benitez, R., Li, Z., Wei, C., Soligalla, R. D., Aizawa, E., Hatanaka, F., Kurita, M., Reddy, P., Ocampo, A., Hishida, T., Sakurai, M., Nemeth, A. N., Nuñez Delicado, E., Campistol, J. M., Magistretti, P., Guillen, P., Rodriguez Esteban, C., ... Izpisua Belmonte, J. C. (2019). Precise in vivo genome editing via single homology arm donor mediated intron-targeting gene integration for genetic disease correction. Cell Research, 29(10), 804-819. https://doi.org/10.1038/s41422-019-0213-0