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

by K Suzuki, Y Tsunekawa, R Benitez, J Wu, J Zhu, E J. Kim, F Hatanaka, M Yamamoto, T Araoka, Z Li, M Kurita, T Hishida, M Li, E Aizawa, S Guo, S Chen, A Goebl, R Soligalla, J Qu, T Jiang, X Fu, M Jafari, Concepcion R Esteban, W Berggren Et Al
Year: 2016


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 Nuñez-Delicado, Pedro Guillen, Josep M. Campistol, Fumio Matsuzaki, Guang-Hui Liu, Pierre Magistretti, Kun Zhang, Edward M. Callaway, Kang Zhang and Juan Carlos Izpisua Belmonte. "In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration​" Nature volume 540, (2016): ​​​144–149 ​​​


​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. 


Genome CRISPR Cas9