The CRISPR system: a new frontier in prokaryotic molecular biology

CRISPR系统：原核分子生物学的新前沿

• 批准号: BB/G011400/1
• 负责人: Malcolm White
• 金额: $ 101.02万
• 依托单位: University of St Andrews
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2009
• 资助国家: 英国
• 起止时间: 2009 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FG011400%2F1
• 关键词: CRISPR; system; new; frontier; prokaryotic

The battle between viruses and the cells they attempt to infect and subvert is one of the main driving forces in molecular evolution. Prokaryotes target and degrade invading DNA using 'restriction enzymes', the discovery of which revolutionised molecular biology. Eukaryotes, including humans, use the RNA interference (RNAi) pathway to target and degrade viral RNA. In the last few years a prokaryotic version of the RNAi pathway, the so-called CAS-CRISPR system, has come to light. Viral sequences are stored in the host genome as 'spacers' flanked by direct repeat sequences (CRISPRs). It is known that host cells incorporate new viral DNA into the CRISPRs, and that this confers immunity against future viral infection. The CRISPR system is envisaged to act in an analogous manner to the RNAi pathway A number of widely conserved 'Cas' proteins (up to 50 per genome) are associated with the CRISPR sequences, and these are thought to be responsible for CRISPR processing, viral defence and the incorporation of new viral sequences. The functions of these proteins are not clearly understood. We have the experience and technological know-how to study all the Cas proteins from the model organism Sulfolobus solfataricus. Cas protein structures will be solved, and Cas protein interactions delineated. The activities of individual Cas proteins and complexes will be determined using a variety of techniques in routine use in our laboratories. The ultimate aim is to acheive a molecular understanding of the CRISPR system. As well as providing fundamental new insights into prokaryotic biology, the work may lead to new methods to manipulate the genomes of useful organisms (synthetic biology) or to treatments for important microbial pathogens.

病毒与它们试图感染和破坏的细胞之间的战斗是分子进化的主要驱动力之一。原核生物使用“限制酶”靶向并降解入侵的DNA，这一发现彻底改变了分子生物学。包括人类在内的真核生物使用RNA干扰（RNAi）途径来靶向和降解病毒RNA。在过去的几年里，RNAi途径的原核版本，即所谓的CAS-CRISPR系统，已经曝光。病毒序列作为“间隔区”储存在宿主基因组中，其侧翼为直接重复序列（CRISPR）。已知宿主细胞将新的病毒DNA整合到CRISPR中，并且这赋予了针对未来病毒感染的免疫力。CRISPR系统被设想以类似于RNAi途径的方式起作用。许多广泛保守的“Cas”蛋白（每个基因组多达50个）与CRISPR序列相关，并且这些被认为负责CRISPR加工、病毒防御和新病毒序列的掺入。这些蛋白质的功能尚不清楚。我们拥有研究模式生物硫磺硫化叶菌（Sulfolobus solfataricus）的所有Cas蛋白的经验和技术知识。Cas蛋白的结构将被解决，Cas蛋白的相互作用划定。单个Cas蛋白和复合物的活性将使用我们实验室常规使用的各种技术来确定。最终目的是从分子水平上理解CRISPR系统。除了为原核生物学提供基本的新见解外，这项工作还可能导致操纵有用生物体基因组的新方法（合成生物学）或重要微生物病原体的治疗方法。

项目成果

CRISPR interference: a structural perspective.

• DOI: 10.1042/bj20130316
• 发表时间: 2013-07-15
• 期刊: The Biochemical journal
• 作者: Reeks J;Naismith JH;White MF
• 通讯作者: White MF

Structure of a dimeric crenarchaeal Cas6 enzyme with an atypical active site for CRISPR RNA processing.

• DOI: 10.1042/bj20130269
• 发表时间: 2013-06-01
• 期刊: The Biochemical journal
• 作者: Reeks J;Sokolowski RD;Graham S;Liu H;Naismith JH;White MF
• 通讯作者: White MF

Structure of the archaeal Cascade subunit Csa5: relating the small subunits of CRISPR effector complexes.

• DOI: 10.4161/rna.23854
• 发表时间: 2013-05
• 期刊: RNA biology
• 影响因子: 4.1
• 作者: Reeks J;Graham S;Anderson L;Liu H;White MF;Naismith JH
• 通讯作者: Naismith JH

Structure and mechanism of the CMR complex for CRISPR-mediated antiviral immunity.

• DOI: 10.1016/j.molcel.2011.12.013
• 发表时间: 2012-02-10
• 期刊: MOLECULAR CELL
• 影响因子: 16
• 作者: Zhang, Jing;Rouillon, Christophe;Kerou, Melina;Reeks, Judith;Brugger, Kim;Graham, Shirley;Reimann, Julia;Cannone, Giuseppe;Liu, Huanting;Albers, Sonja-Verena;Naismith, James H.;Spagnolo, Laura;White, Malcolm F.
• 通讯作者: White, Malcolm F.

The CRISPR associated protein Cas4 Is a 5' to 3' DNA exonuclease with an iron-sulfur cluster.

• DOI: 10.1371/journal.pone.0047232
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Zhang J;Kasciukovic T;White MF
• 通讯作者: White MF