In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration

Autor: Euiseok J. Kim, Tomoaki Hishida, Fumiyuki Hatanaka, Estrella Núñez-Delicado, Edward M. Callaway, Shicheng Guo, Pierre J. Magistretti, Kang Zhang, Tingshuai Jiang, Reyna Hernández-Benítez, Kun Zhang, Jie Zhu, Concepcion Rodriguez Esteban, Masakazu Kurita, W. Travis Berggren, Jun Wu, Mo Li, Song Chen, Pedro Guillen, Yuji Tsunekawa, April Goebl, Toshikazu Araoka, Jing Qu, Rupa Devi Soligalla, Jeronimo Lajara, Maryam Jafari, Guang-Hui Liu, Keiichiro Suzuki, Fumio Matsuzaki, Mako Yamamoto, Zhe Li, Xin Fu, Juan Carlos Izpisua Belmonte, Emi Aizawa, Josep M. Campistol
Rok vydání: 2016
Předmět:
Zdroj: Nature. 540:144-149
ISSN: 1476-4687
0028-0836
Popis: 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.
Databáze: OpenAIRE
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