The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli.
The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli.
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DOI：
10.1093/nar/gkr606
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发表时间：
2011-11
影响因子：
14.9
通讯作者：
Siksnys V
中科院分区：
文献类型：
--
作者：
Sapranauskas R;Gasiunas G;Fremaux C;Barrangou R;Horvath P;Siksnys V
CRISPR/CAS适应性免疫系统可抵抗古细菌和细菌中的噬菌体和质粒。 CRISPR基因座从入侵的遗传因素中整合了短的DNA序列，这些遗传元素在随后暴露于匹配的核酸时提供了小的RNA介导的干扰。在嗜热链球菌中，以前证明，CRISPR1/CAS系统可以通过在暴露于这些异物遗传元件后整合新的间隔物来提供对噬菌体和质粒的适应性免疫，这些遗传元素随后指导了侵入性同源DNA序列的特定裂解。在这里，我们表明，嗜热链球菌CRISPR3/CAS系统可以转移到大肠杆菌中，并为质粒转化和噬菌体感染提供异源保护。我们表明干扰是序列特异性的，并且附近或原始间隔基邻近基序（PAM）中的突变使质粒可以逃脱CIRSPR编码的免疫力。我们还确定CAS9是CRIS编码干扰所必需的唯一CAS基因。此外，突变分析表明，干扰依赖于Cas9 Mcra/hnh-和ruvc/rnaseh-motifs。总的来说，我们的结果表明，活动的CRISPR/CAS系统可以在遥远的属上转移，并提供针对浸润性核酸的异源干扰。可以利用这是为了在噬菌体攻击方面发展菌株，并且更安全的生物摄入和传播质粒编码的不良遗传元件的可能性较小。
The CRISPR/Cas adaptive immune system provides resistance against phages and plasmids in Archaea and Bacteria. CRISPR loci integrate short DNA sequences from invading genetic elements that provide small RNA-mediated interference in subsequent exposure to matching nucleic acids. In Streptococcus thermophilus, it was previously shown that the CRISPR1/Cas system can provide adaptive immunity against phages and plasmids by integrating novel spacers following exposure to these foreign genetic elements that subsequently direct the specific cleavage of invasive homologous DNA sequences. Here, we show that the S. thermophilus CRISPR3/Cas system can be transferred into Escherichia coli and provide heterologous protection against plasmid transformation and phage infection. We show that interference is sequence-specific, and that mutations in the vicinity or within the proto-spacer adjacent motif (PAM) allow plasmids to escape CRISPR-encoded immunity. We also establish that cas9 is the sole cas gene necessary for CRISPR-encoded interference. Furthermore, mutation analysis revealed that interference relies on the Cas9 McrA/HNH- and RuvC/RNaseH-motifs. Altogether, our results show that active CRISPR/Cas systems can be transferred across distant genera and provide heterologous interference against invasive nucleic acids. This can be leveraged to develop strains more robust against phage attack, and safer organisms less likely to uptake and disseminate plasmid-encoded undesirable genetic elements.
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DOI：
10.1126/science.1165771
发表时间：
2008-12-19
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Marraffini LA;Sontheimer EJ
通讯作者：
Sontheimer EJ
影响因子：
3.2
作者：
Deveau, Helene;Barrangou, Rodolphe;Moineau, Sylvain
通讯作者：
Moineau, Sylvain
影响因子：
3.9
作者：
Mojica, FJM;Díez-Villaseñor, C;Soria, E
通讯作者：
Soria, E
新型克隆载体的构建和特性分析 2. 多用途克隆系统
影响因子：
3.5
作者：
BOLIVAR, F;RODRIGUEZ, RL;FALKOW, S
通讯作者：
FALKOW, S
DOI：
10.1126/science.1159689
发表时间：
2008-08-15
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Brouns SJ;Jore MM;Lundgren M;Westra ER;Slijkhuis RJ;Snijders AP;Dickman MJ;Makarova KS;Koonin EV;van der Oost J
通讯作者：
van der Oost J