The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA
The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA
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DOI：
10.1038/nature09523
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发表时间：
2010-11-04
期刊：
影响因子：
64.8
通讯作者：
Moineau, Sylvain
中科院分区：
文献类型：
--
作者：
Garneau, Josiane E.;Dupuis, Marie-Eve;Moineau, Sylvain
细菌和病原菌已经发展了几种防御外来核酸如病毒基因组和质粒的策略。其中，成簇的规则间隔的短回文重复序列（CRISPR）基因座与cas（CRISPR相关）基因一起形成CRISPR/Cas免疫系统，其涉及由称为间隔子的短DNA片段分离的部分回文重复序列，其从染色体外元件获得。最近证明，这些可变基因座可以并入来自感染噬菌体的间隔区，然后以序列特异性方式提供针对随后的噬菌体感染的免疫力。在这里，我们表明，嗜热链球菌CRISPR 1/Cas系统也可以自然地从含有耐药性基因的自我复制质粒中获得间隔区，导致质粒丢失。获得的与耐药基因相匹配的间隔区提供了一种新的手段，可以自然选择不能吸收和传播这些基因的细菌。我们还提供了体内证据，证明CRISPR 1/Cas系统在特定位点特异性切割原型间隔区内的质粒和噬菌体双链DNA。我们的数据表明，CRISPR/Cas免疫系统非常适合快速切割入侵DNA，并有潜力开发出更安全的微生物菌株。
Bacteria and Archaea have developed several defence strategies against foreign nucleic acids such as viral genomes and plasmids. Among them, clustered regularly interspaced short palindromic repeats (CRISPR) loci together with cas (CRISPR-associated) genes form the CRISPR/Cas immune system, which involves partially palindromic repeats separated by short stretches of DNA called spacers, acquired from extrachromosomal elements. It was recently demonstrated that these variable loci can incorporate spacers from infecting bacteriophages and then provide immunity against subsequent bacteriophage infections in a sequence-specific manner. Here we show that the Streptococcus thermophilus CRISPR1/Cas system can also naturally acquire spacers from a self-replicating plasmid containing an antibiotic-resistance gene, leading to plasmid loss. Acquired spacers that match antibiotic-resistance genes provide a novel means to naturally select bacteria that cannot uptake and disseminate such genes. We also provide in vivo evidence that the CRISPR1/Cas system specifically cleaves plasmid and bacteriophage double-stranded DNA within the proto-spacer, at specific sites. Our data show that the CRISPR/Cas immune system is remarkably adapted to cleave invading DNA rapidly and has the potential for exploitation to generate safer microbial strains.