In Vivo Genome Editing Via CRISPR/Cas9 Mediated Homology-independent Targeted Integration

Overview

Journal Nature

Specialty Science

Date 2016 Nov 17

PMID 27851729

Citations 577

Authors

Keiichiro Suzuki

Yuji Tsunekawa

Reyna Hernandez-Benitez

Jun Wu

Jie Zhu

Euiseok J Kim

Fumiyuki Hatanaka

Mako Yamamoto

Toshikazu Araoka

Zhe Li

Masakazu Kurita

Tomoaki Hishida

Mo Li

Emi Aizawa

Shicheng Guo

Song Chen

April Goebl

Rupa Devi Soligalla

Jing Qu

Tingshuai Jiang

Xin Fu

Maryam Jafari

Concepcion Rodriguez Esteban

W Travis Berggren

Jeronimo Lajara

Estrella Nunez-Delicado

Pedro Guillen

Josep M Campistol

Fumio Matsuzaki

Guang-Hui Liu

Pierre Magistretti

Kun Zhang

Edward M Callaway

Kang Zhang

Juan Carlos Izpisua Belmonte

Affiliations

Soon will be listed here.

Abstract

Targeted genome editing via engineered nucleases is an exciting area of biomedical research and holds potential for clinical applications. Despite rapid advances in the field, in vivo targeted transgene integration is still infeasible because current tools are inefficient, especially for non-dividing cells, which compose most adult tissues. This poses a barrier for uncovering fundamental biological principles and developing treatments for a broad range of genetic disorders. Based on clustered regularly interspaced short palindromic repeat/Cas9 (CRISPR/Cas9) technology, here we devise a homology-independent targeted integration (HITI) strategy, which allows for robust DNA knock-in in both dividing and non-dividing cells in vitro and, more importantly, in vivo (for example, in neurons of postnatal mammals). As a proof of concept of its therapeutic potential, we demonstrate the efficacy of HITI in improving visual function using a rat model of the retinal degeneration condition retinitis pigmentosa. The HITI method presented here establishes new avenues for basic research and targeted gene therapies.

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