利用嗜热泉古菌 Argonaute 进行 DNA 引导的基因组编辑。
DNA-guided genome editing using the Natronobacterium gregoryi Argonaute.
机构信息
Department of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhuang, Hebei, China.
Department of Physiology, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
出版信息
Nat Biotechnol. 2016 Jul;34(7):768-73. doi: 10.1038/nbt.3547. Epub 2016 May 2.
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 ∼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.
RNA 指导的内切酶 Cas9 使基因组编辑成为一种广泛应用的技术。类似于 Cas9，Argonaute 蛋白家族的内切酶也使用寡核苷酸作为向导来降解入侵的基因组。在这里，我们报告说，来源于 Natronobacterium gregoryi 的 Argonaute（NgAgo）是一种适用于人类细胞基因组编辑的 DNA 指导内切酶。NgAgo 结合 5' 磷酸化的单链引导 DNA（gDNA）约 24 个核苷酸，当加载 gDNA 时，可有效地在特定位点产生 DNA 双链断裂。与 Cas9 不同，NgAgo-gDNA 系统不需要原间隔基序 (PAM)，并且初步特征表明其对引导物-靶标错配的容忍度低，并且在编辑富含 (G+C) 的基因组靶标时效率高。
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