In vivo genome editing restores haemostasis in a mouse model of haemophilia.
In vivo genome editing restores haemostasis in a mouse model of haemophilia.
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DOI：
10.1038/nature10177
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发表时间：
2011-06-26
期刊：
影响因子：
64.8
通讯作者：
High KA
中科院分区：
文献类型：
--
作者：
Li H;Haurigot V;Doyon Y;Li T;Wong SY;Bhagwat AS;Malani N;Anguela XM;Sharma R;Ivanciu L;Murphy SL;Finn JD;Khazi FR;Zhou S;Paschon DE;Rebar EJ;Bushman FD;Gregory PD;Holmes MC;High KA
编辑人类基因组以纠正致病突变是治疗遗传性疾病的一种有前途的方法。基因组编辑通过实现突变基因的原位校正来改进简单的基因替换策略，从而在内源性调控元件的控制下恢复正常的基因功能，并降低与随机插入基因组相关的风险。基因特异性靶向历来仅限于小鼠胚胎干细胞。锌指核酸酶（ZFN）的发展已经允许在转化细胞和原代细胞中进行有效的基因组编辑，这些转化细胞和原代细胞先前被认为难以进行这种遗传操作。在体外，ZFN已经显示出通过在靶基因座处诱导位点特异性双链断裂（DSB），但目前尚不清楚ZFN是否能在体内诱导DSB并刺激具有临床意义的基因组编辑。在这里，我们表明，ZFN是能够有效地诱导DSB时，直接交付给小鼠肝脏，并与适当设计的基因靶向载体共同交付时，他们可以刺激基因置换，通过同源定向和同源性无关的靶向基因插入ZFN指定的基因座。达到的基因靶向水平足以纠正血友病B小鼠模型中的凝血时间延长，并在诱导肝再生后保持持续。因此，ZFN驱动的基因校正可以在体内进行，提高了基因组编辑作为治疗遗传疾病的可行策略的可能性。
Editing of the human genome to correct disease-causing mutations is a promising approach for the treatment of genetic disorders. Genome editing improves on simple gene-replacement strategies by effectingin situcorrection of a mutant gene, thus restoring normal gene function under the control of endogenous regulatory elements and reducing risks associated with random insertion into the genome. Gene-specific targeting has historically been limited to mouse embryonic stem cells. The development of zinc finger nucleases (ZFNs) has permitted efficient genome editing in transformed and primary cells that were previously thought to be intractable to such genetic manipulation.In vitro, ZFNs have been shown to promote efficient genome editing via homology-directed repair by inducing a site-specific double-strand break (DSB) at a target locus,,, but it is unclear whether ZFNs can induce DSBs and stimulate genome editing at a clinically meaningful levelin vivo. Here we show that ZFNs are able to induce DSBs efficiently when delivered directly to mouse liver and that, when co-delivered with an appropriately designed gene-targeting vector, they can stimulate gene replacement through both homology-directed and homology-independent targeted gene insertion at the ZFN-specified locus. The level of gene targeting achieved was sufficient to correct the prolonged clotting times in a mouse model of haemophilia B, and remained persistent after induced liver regeneration. Thus, ZFN-driven gene correction can be achievedin vivo, raising the possibility of genome editing as a viable strategy for the treatment of genetic disease.