相关实验视频
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Precise Phage Mutagenesis with NgTET-Assisted CRISPR-Cas Systems
Published on: October 14, 2025
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通过菌体蛋白抑制CRISPR-Cas9
Benjamin J Rauch1, Melanie R Silvis2, Judd F Hultquist3
1Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA; Quantitative Biosciences Institute, QBI, University of California, San Francisco, San Francisco, CA 94158, USA.
Cell
|January 3, 2017
概括
研究人员发现了天然的CRISPR-Cas9抑制剂, 这些蛋白质阻断了CRISPR-Cas9基因组编辑,为控制基因编辑技术提供了新的工具.
科学领域:
- 微生物学
- 分子生物学
- 遗传学
背景情况:
- 细菌CRISPR-Cas系统使用RNA导向核酶提供对菌体的适应性免疫力.
- 菌体已经进化出抗CRISPR蛋白来抑制1类CRISPR-Cas系统.
- 对于广泛使用的CRISPR- Cas9系统,没有已知的天然抑制剂.
研究的目的:
- 确定抑制CRISPR-Cas9系统的蛋白质.
- 研究CRISPR-Cas9失活的自然机制.
主要方法:
- 对含有cas9的细菌基因组进行生物信息搜索,以寻找共存的CRISPR间隔器和标.
- 来自Listeria monocytogenes的潜在抑制蛋白的识别和表征.
- 用大肠杆菌和人类细胞进行功能测试以确认抑制活性.
主要成果:
- 发现了四种独特的II-ACRISPR-Cas9抑制蛋白质,这些蛋白质是由Listeria monocytogenes的prophages编码的.
- 超过50%的Listeria monocytogenes菌株含有cas9,它们至少含有一种传染体编码的抑制剂.
- 两种已确定的抑制剂有效地阻断了Streptococcus pyogenes在细菌和人类细胞中的Cas9活性.
结论:
- 天然的CRISPR-Cas9抑制剂,称为抗CRISPRs,存在并且在Listeria monocytogenes中广泛存在.
- 这些抗CRISPR蛋白代表了一类新的Cas9抑制剂.
- 发现的抗CRISPR可以作为调节CRISPR-Cas9基因组编辑应用的工具.
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