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Cell Res. 2019 Aug 23. doi: 10.1038/s41422-019-0213-0. [Epub ahead of print]

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

Author information

1
Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, CA, 92037, USA. ksuzuki@chem.es.osaka-u.ac.jp.
2
Institute for Advanced Co-Creation Studies, Osaka University, Osaka, 560-8531, Japan. ksuzuki@chem.es.osaka-u.ac.jp.
3
Graduate School of Engineering Science, Osaka University, Osaka, 560-8531, Japan. ksuzuki@chem.es.osaka-u.ac.jp.
4
Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, CA, 92037, USA.
5
Bioengineering, University of California, San Diego, 9500 Gilman Drive, MC0412, La Jolla, CA, 92093-0412, USA.
6
Universidad Catolica, San Antonio de Murcia, Campus de los Jeronimos, 135, Guadalupe, 30107, Spain.
7
Graduate School of Engineering Science, Osaka University, Osaka, 560-8531, Japan.
8
Hospital Clinic of Barcelona, Carrer Villarroel, 170, 08036, Barcelona, Spain.
9
King Abdullah University of Science and Technology (KAUST), Thuwal, 23955-6900, Saudi Arabia.
10
Fundacion Dr. Pedro Guillen, Clinica CEMTRO, Avenida Ventisquero de la Condesa, 4228035, Madrid, Spain.
11
BGI-Shenzhen, Shenzhen, 518083, China.
12
Guangdong Provincial Key Laboratory of Genome Read and Write, Shenzhen, 518120, China.
13
Guangdong Provincial Academician Workstation of BGI Synthetic Genomics, BGI-Shenzhen, Guangdong, China.
14
Shenzhen Engineering Laboratory for Innovative Molecular Diagnostics, Shenzhen, 518120, China.
15
National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
16
Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Instituto Universitario de Oncología (IUOPA), Universidad de Oviedo, Oviedo, Spain.
17
Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA.
18
Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA.
19
Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, CA, 92037, USA. belmonte@salk.edu.

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.

PMID:
31444470
DOI:
10.1038/s41422-019-0213-0

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