<br>CRISPR-Cas9 的温度响应性竞争性抑制。,Molecular Cell

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CRISPR-Cas9 的温度响应性竞争性抑制。
Molecular Cell ( IF 14.5 ) Pub Date : 2018-12-27 , DOI: 10.1016/j.molcel.2018.11.016
Fuguo Jiang ₁ , Jun-Jie Liu ₂ , Beatriz A Osuna ₃ , Michael Xu ₄ , Joel D Berry ₅ , Benjamin J Rauch ₅ , Eva Nogales ₆ , Joseph Bondy-Denomy ₅ , Jennifer A Doudna ₇

Affiliation

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA 94720, USA.
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; MBIB Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA.
Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720, USA.
Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA; California Institute for Quantitative Biosciences, University of California, San Francisco, San Francisco, CA 94143, USA.
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720, USA; Howard Hughes Medical Institute, Berkeley; MBIB Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA 94720, USA; Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720, USA; Innovative Genomics Institute, Berkeley, CA 94704, USA; Howard Hughes Medical Institute, Berkeley; MBIB Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA; Gladstone Institutes, San Francisco, CA 94158, USA.

CRISPR-Cas 免疫系统利用 RNA 引导的核酸酶来保护细菌免受噬菌体感染。噬菌体反过来又进化出了抑制性“抗 CRISPR”(Acr) 蛋白，包括六种抑制剂 (AcrIIA1-AcrIIA6)，它们可以阻止 II-A 型 CRISPR-Cas9 酶进行 DNA 切割和基因组编辑。我们在此表明，AcrIIA2 及其更有效的同源物 AcrIIA2b 通过封闭 DNA 结合所需的蛋白质残基来阻止 Cas9 与 DNA 结合。冷冻电镜测定的 AcrIIA2 或 AcrIIA2b 与化脓性链球菌 Cas9 结合的结构揭示了一种与其他已知 Acrs 不同的 DNA 结合竞争性抑制模式。 AcrIIA2 和 AcrIIA2b 对 Cas9 抑制的温度依赖性差异源于 Cas9 上抑制剂结构和局部抑制剂结合环境的差异。这些发现扩展了在时间、空间和条件上调节 CRISPR-Cas9 基因组编辑的天然工具箱。

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更新日期：2018-12-27

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