Cas9 function and host genome sampling in Type II-A CRISPR-Cas adaptation.
Cas9 function and host genome sampling in Type II-A CRISPR-Cas adaptation.
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DOI：
10.1101/gad.257550.114
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发表时间：
2015-02-15
影响因子：
10.5
通讯作者：
Terns MP
中科院分区：
文献类型：
--
作者：
Wei Y;Terns RM;Terns MP
Wei等人发现，Cas9，以前被鉴定为负责最终入侵者破坏的核酸酶，也是嗜热链球菌适应所必需的。Cas9核酸酶活性是适应性的。Wei等人还揭示了通过CRISPR-Cas系统对自靶向宿主基因组序列的广泛、无偏见的获取，该系统在存在主动靶破坏的情况下被掩蔽。为了获得识别和破坏病毒和质粒入侵者的能力，原核CRISPR-Cas系统捕获宿主CRISPR基因座内的DNA片段。我们的研究结果表明，嗜热链球菌中II-A型CRISPR-Cas系统的适应过程需要Cas 1，Cas 2和Csn 2。令人惊讶的是，我们发现Cas9，以前被确定为负责最终入侵者破坏的核酸酶，也是适应所必需的。Cas9核酸酶活性是适应性的。此外，我们的研究揭示了CRISPR-Cas系统对自靶向宿主基因组序列的广泛、公正的获取，该系统在存在主动靶向破坏的情况下被掩盖。
Wei et al. found that Cas9, previously identified as the nuclease responsible for ultimate invader destruction, is also essential for adaptation in Streptococcus thermophilus. Cas9 nuclease activity is dispensable for adaptation. Wei et al. also revealed extensive, unbiased acquisition of the self-targeting host genome sequence by the CRISPR–Cas system that is masked in the presence of active target destruction. To acquire the ability to recognize and destroy virus and plasmid invaders, prokaryotic CRISPR–Cas systems capture fragments of DNA within the host CRISPR locus. Our results indicate that the process of adaptation by a Type II-A CRISPR–Cas system in Streptococcus thermophilus requires Cas1, Cas2, and Csn2. Surprisingly, we found that Cas9, previously identified as the nuclease responsible for ultimate invader destruction, is also essential for adaptation. Cas9 nuclease activity is dispensable for adaptation. In addition, our studies revealed extensive, unbiased acquisition of the self-targeting host genome sequence by the CRISPR–Cas system that is masked in the presence of active target destruction.
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通过反式编码的小 RNA 和宿主因子 RNase III 进行 CRISPR RNA 成熟。
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