Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland
Institute of Molecular Systems Biology, Eidgenössische Technische Hochschule Zurich, Zurich, Switzerland.
Life Sci Alliance. 2019 Jun 13;2(3). doi: 10.26508/lsa.201800267. Print 2019 Jun.
CRISPR-Cas9-based genome editing has transformed the life sciences, enabling virtually unlimited genetic manipulation of genomes: The RNA-guided Cas9 endonuclease cuts DNA at a specific target sequence and the resulting double-strand breaks are mended by one of the intrinsic cellular repair pathways. Imprecise double-strand repair will introduce random mutations such as indels or point mutations, whereas precise editing will restore or specifically edit the locus as mandated by an endogenous or exogenously provided template. Recent studies indicate that CRISPR-induced DNA cuts may also result in the exchange of genetic information between homologous chromosome arms. However, conclusive data of such recombination events in higher eukaryotes are lacking. Here, we show that in , the detected Cas9-mediated editing events frequently resulted in germline-transmitted exchange of chromosome arms-often without indels. These findings demonstrate the feasibility of using the system for generating recombinants and also highlight an unforeseen risk of using CRISPR-Cas9 for therapeutic intervention.
基于 CRISPR-Cas9 的基因组编辑技术改变了生命科学，使得对基因组进行几乎无限的遗传操作成为可能：RNA 引导的 Cas9 内切酶在特定的靶序列处切割 DNA，而产生的双链断裂则通过内在的细胞修复途径之一进行修复。非精确的双链修复会引入随机突变，如插入缺失或点突变，而精确的编辑则会根据内源性或外源性模板的要求来恢复或特异性地编辑基因座。最近的研究表明，CRISPR 诱导的 DNA 切割也可能导致同源染色体臂之间遗传信息的交换。然而，在高等真核生物中缺乏此类重组事件的明确数据。在这里，我们表明在 中，检测到的 Cas9 介导的编辑事件经常导致染色体臂的种系传递交换-通常没有插入缺失。这些发现证明了该系统用于产生重组体的可行性，并且还突出了使用 CRISPR-Cas9 进行治疗干预的意外风险。
