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New CRISPR-Cas systems from uncultivated microbes

机译：来自未培养微生物的新CRISPR-Cas系统

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摘要

CRISPR-Cas systems provide microbes with adaptive immunity by employing short DNA sequences, termed spacers, that guide Cas proteins to cleave foreign DNA(1,2). Class 2 CRISPR-Cas systems are streamlined versions, in which a single RNA-bound Cas protein recognizes and cleaves target sequences(3,4). The programmable nature of these minimal systems has enabled researchers to repurpose them into a versatile technology that is broadly revolutionizing biological and clinical research(5). However, current CRISPR-Cas technologies are based solely on systems from isolated bacteria, leaving the vast majority of enzymes from organisms that have not been cultured untapped. Metagenomics, the sequencing of DNA extracted directly from natural microbial communities, provides access to the genetic material of a huge array of uncultivated organisms(6,7). Here, using genome-resolved metagenomics, we identify a number of CRISPR-Cas systems, including the first reported Cas9 in the archaeal domain of life, to our knowledge. This divergent Cas9 protein was found in little studied nanoarchaea as part of an active CRISPR-Cas system. In bacteria, we discovered two previously unknown systems, CRISPR-CasX and CRISPR-CasY, which are among the most compact systems yet discovered. Notably, all required functional components were identified by metagenomics, enabling validation of robust in vivo RNA-guided DNA interference activity in Escherichia coli. Interrogation of environmental microbial communities combined with in vivo experiments allows us to access an unprecedented diversity of genomes, the content of which will expand the repertoire of microbe-based biotechnologies.
机译：CRISPR-Cas系统利用短的DNA序列（称为间隔子）为微生物提供适应性免疫，该序列指导Cas蛋白裂解外源DNA（1,2）。 2级CRISPR-Cas系统是简化版本，其中单个RNA结合的Cas蛋白识别并切割靶序列（3,4）。这些最小系统的可编程性质使研究人员能够将它们重新调整为一种用途广泛的技术，从而彻底改变生物学和临床研究（5）。但是，当前的CRISPR-Cas技术仅基于分离细菌的系统，而来自尚未培养的生物体的绝大多数酶仍未开发。元基因组学是对直接从天然微生物群落中提取的DNA进行测序的方法，它提供了获取大量未经培养的生物的遗传物质的途径（6,7）。在这里，据我们所知，使用基因组解析的宏基因组学，我们确定了许多CRISPR-Cas系统，包括生命古细菌域中首次报道的Cas9。这种差异化的Cas9蛋白是作为主动CRISPR-Cas系统的一部分在很少研究的纳古菌中发现的。在细菌中，我们发现了两个以前未知的系统，CRISPR-CasX和CRISPR-CasY，它们是迄今为止发现的最紧凑的系统。值得注意的是，所有必需的功能组件均通过宏基因组学进行了鉴定，从而能够验证大肠杆菌中强大的体内RNA引导的DNA干扰活性。对环境微生物群落的询问与体内实验相结合，使我们能够访问前所未有的基因组多样性，其内容将扩大基于微生物的生物技术的种类。

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  • 来源
    《Nature》 |2017年第7640期|237-241|共5页
  • 作者

    Burstein David; Harrington Lucas B.; Strutt Steven C.; Probst Alexander J.; Anantharaman Karthik; Thomas Brian C.; Doudna Jennifer A.; Banfield Jillian F.;

  • 作者单位

    Univ Calif Berkeley, Dept Earth & Planetary Sci, Berkeley, CA 94720 USA;

    Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA;

    Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA;

    Univ Calif Berkeley, Dept Earth & Planetary Sci, Berkeley, CA 94720 USA;

    Univ Calif Berkeley, Dept Earth & Planetary Sci, Berkeley, CA 94720 USA;

    Univ Calif Berkeley, Dept Earth & Planetary Sci, Berkeley, CA 94720 USA;

    Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA|Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA|Univ Calif Berkeley, Howard Hughes Med Inst, Berkeley, CA 94720 USA|Univ Calif Berkeley, Innovat Genom Initiat, Berkeley, CA 94720 USA|Lawrence Berkeley Natl Lab, MBIB Div, Berkeley, CA 94720 USA;

    Univ Calif Berkeley, Dept Earth & Planetary Sci, Berkeley, CA 94720 USA|Univ Calif Berkeley, Dept Environm Sci Policy & Management, Berkeley, CA 94720 USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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