作者
Keiichiro Suzuki,Yuji Tsunekawa,Reyna Hernández-Benítez,Jun Wu,Jie Zhu,Euiseok J. Kim,Fumiyuki Hatanaka,Masaaki Yamamoto,Toshikazu Araoka,Zhe Li,Masanori Kurita,Tomoaki Hishida,Li Mo,Emi Aizawa,Shicheng Guo,Song Chen,April Goebl,Rupa Devi Soligalla,Jing Qu,Tongtong Jiang,Xin Fu,Maryam Jafari,Concepción Rodrı́guez Esteban,W. Travis Berggren,Jerónimo Lajara,Estrella Núñez‐Delicado,Pedro Guillén,Josep M. Campistol,Fumio Matsuzaki,Guanghui Liu,Pierre J. Magistretti,Kun Zhang,Edward M. Callaway,Kang Zhang,Juan Carlos Izpisúa Belmonte
摘要
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.