DNA-guided genome editing using the Natronobacterium gregoryi Argonaute (Publication with Expression of Concern. See vol. 35, pg. 481, 2017) (Retracted article. See vol. 35, pg. 797, 2017)
DNA-guided genome editing using the Natronobacterium gregoryi Argonaute (Publication with Expression of Concern. See vol. 35, pg. 481, 2017) (Retracted article. See vol. 35, pg. 797, 2017)
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DOI：
10.1038/nbt.3547
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发表时间：
2016-07-01
影响因子：
46.9
通讯作者：
Han, Chunyu
中科院分区：
文献类型：
--
作者：
Gao, Feng;Shen, Xiao Z.;Han, Chunyu
RNA引导的内切酶Cas9使基因组编辑成为一项广泛使用的技术。与Cas9类似，ArgAerte蛋白家族的内切酶也使用寡核苷酸作为降解侵袭性基因组的指南。在此，我们报道了NgAgo是一种DNA引导的核酸内切酶，适用于人类细胞的基因组编辑。NgAgo与类似于24个核苷酸的5‘磷酸化单链引导DNA(GDNA)结合，当加载gDNA时有效地产生位点特异性DNA双链断裂。与Cas9一样，NgAgo-gDNA系统不需要ProtSpacer相邻基序(PAM)，初步特征表明对引导靶错配的耐受性低，编辑富含(G+C)的基因组靶的效率高。
The RNA-guided endonuclease Cas9 has made genome editing a widely accessible technique. Similar to Cas9, endonucleases from the Argonaute protein family also use oligonucleotides as guides to degrade invasive genomes. Here we report that the Natronobacterium gregoryi Argonaute (NgAgo) is a DNA-guided endonuclease suitable for genome editing in human cells. NgAgo binds 5' phosphorylated single-stranded guide DNA (gDNA) of similar to 24 nucleotides, efficiently creates site-specific DNA double-strand breaks when loaded with the gDNA. The NgAgo-gDNA system does not require a protospacer-adjacent motif (PAM), as does Cas9, and preliminary characterization suggests a low tolerance to guide-target mismatches and high efficiency in editing (G+C)-rich genomic targets.