Correction of a pathogenic gene mutation in human embryos (Publication with Expression of Concern)
Correction of a pathogenic gene mutation in human embryos (Publication with Expression of Concern)
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DOI：
10.1038/nature23305
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发表时间：
2017-08-24
期刊：
影响因子：
64.8
通讯作者：
Mitalipov, Shoukhrat
中科院分区：
文献类型：
--
作者：
Ma, Hong;Marti-Gutierrez, Nuria;Mitalipov, Shoukhrat
基因组编辑具有靶向纠正种系突变的潜力。在这里，我们描述了通过激活内源性、生殖系特异性DNA修复反应，以精确的基于CRISPR-Cas9的靶向准确性和高同源性定向修复效率来校正人类植入前胚胎中的杂合MYBPC 3突变。诱导的双链断裂（DSB）在突变的父系等位基因主要修复使用同源野生型母系基因，而不是合成的DNA模板。通过调节诱导DSB的细胞周期阶段，我们能够避免切割胚胎中的镶嵌现象，并获得高产量的携带野生型MYBPC 3基因的纯合胚胎，而没有脱靶突变的证据。所提出的方法的效率，准确性和安全性表明，它有可能用于纠正人类胚胎中的遗传突变，通过补充植入前遗传学诊断。然而，在临床应用之前仍有许多问题需要考虑，包括该技术与其他杂合突变的重现性。
Genome editing has potential for the targeted correction of germline mutations. Here we describe the correction of the heterozygous MYBPC3 mutation in human preimplantation embryos with precise CRISPR-Cas9-based targeting accuracy and high homology-directed repair efficiency by activating an endogenous, germline-specific DNA repair response. Induced double-strand breaks (DSBs) at the mutant paternal allele were predominantly repaired using the homologous wild-type maternal gene instead of a synthetic DNA template. By modulating the cell cycle stage at which the DSB was induced, we were able to avoid mosaicism in cleaving embryos and achieve a high yield of homozygous embryos carrying the wild-type MYBPC3 gene without evidence of off-target mutations. The efficiency, accuracy and safety of the approach presented suggest that it has potential to be used for the correction of heritable mutations in human embryos by complementing preimplantation genetic diagnosis. However, much remains to be considered before clinical applications, including the reproducibility of the technique with other heterozygous mutations.