In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration.
In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration.
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通过 CRISPR/Cas9 介导的同源独立靶向整合进行体内基因组编辑
DOI：
10.1038/nature20565
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发表时间：
2016-12-01
期刊：
影响因子：
64.8
通讯作者：
Belmonte JC
中科院分区：
文献类型：
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作者：
Suzuki K;Tsunekawa Y;Hernandez-Benitez R;Wu J;Zhu J;Kim EJ;Hatanaka F;Yamamoto M;Araoka T;Li Z;Kurita M;Hishida T;Li M;Aizawa E;Guo S;Chen S;Goebl A;Soligalla RD;Qu J;Jiang T;Fu X;Jafari M;Esteban CR;Berggren WT;Lajara J;Nuñez-Delicado E;Guillen P;Campistol JM;Matsuzaki F;Liu GH;Magistretti P;Zhang K;Callaway EM;Zhang K;Belmonte JC
通过工程核酸酶进行靶向基因组编辑是生物医学研究的一个令人兴奋的领域，具有临床应用的潜力。尽管该领域进展迅速，但体内靶向转基因整合仍然不可行，因为目前的工具效率低下，特别是对于构成大多数成人组织的非分裂细胞。这对揭示基本生物学原理和开发广泛遗传疾病的治疗方法构成了障碍。基于聚类规则间隔短回文重复/Cas9 （CRISPR/Cas9）技术，我们设计了一种同源无关的靶向整合（HITI）策略，该策略允许在体外分裂和非分裂细胞中进行强大的DNA敲入，更重要的是，在体内（例如，在出生后哺乳动物的神经元中）。为了证明其治疗潜力的概念，我们用视网膜变性条件视网膜色素变性的大鼠模型证明了HITI在改善视觉功能方面的功效。本文提出的HITI方法为基础研究和靶向基因治疗开辟了新的途径。
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.