CRISPR Capture, Destroy, and Counter-Attack Mechanisms

项目摘要 CRISPR-CAS适应性免疫系统被原核生物用来保护它们免受潜在的 由病毒引起的致命感染。作为一种防御策略，某些病毒会产生抗CRISPR(ACR)蛋白 能够灭活CRISPR介导的抗病毒免疫。不同的CRISPR-CAS系统通过以下方式运行 捕获宿主基因组CRISPR基因座内的短病毒序列并使用表达的crRNAs和 与病毒识别和破坏相关的Cas核酸酶。病毒ACRS的特性和功能 颠覆独特的CRISPR-CAS系统才刚刚揭开面纱。我们的主要目标是 研究计划是：1)了解两种寄主CRISPR-CAS的分子基础 病毒的免疫途径和抗CRISPR活性以及2)利用所获得的知识来贡献和 改进基于CRISPR的应用程序。利用分子、基因、 结构和生化方法，凭借一支出色的合作者团队做出的宝贵贡献，我们 将继续致力于剖析调控不同CRISPR-CA的途径和描绘分子机制 系统。此外，我们还将深入了解病毒使用ACR介导的机制来阻止 CRISPR豁免权。基于CRISPR的创新工具和技术在以下方面提供了变革性的进步 基础和医学研究。我们通过研究获得的知识将直接有助于正在进行的努力 旨在利用各种CRISPR-CAS系统作为强大的研究工具(例如，用于基因组编辑和 控制基因表达，选择性地检测和靶向人类病毒和细菌病原体，以及 限制抗生素耐药性的传播)。此外，了解不同组织的具体行动模式 ACRS是建立ACRS作为选择性和安全利用的重要新研究工具所必需的 不同的基于CRISPR的技术。

Project Summary CRISPR-Cas adaptive immune systems are deployed by prokaryotic organisms to protect them from potential lethal infection by viruses. As a counter-defense strategy, certain viruses produce anti-CRISPR (ACR) proteins capable of inactivating CRISPR-mediated, anti-viral immunity. Diverse CRISPR-Cas systems function by capturing short viral sequences within the CRISPR locus of the host genome and using expressed crRNAs and associated Cas nucleases for viral recognition and destruction. The identities and functions of viral ACRs in subverting distinct CRISPR-Cas systems are only just now being revealed. The primary objectives of our research program are: 1) to understand the molecular basis of mechanisms underlying both host CRISPR-Cas immune pathways and anti-CRISPR activities of viruses and 2) to exploit knowledge gained to contribute and improve CRISPR-based applications. Using an established and powerful combination of molecular, genetic, structural and biochemical approaches, with invaluable contributions from a team of superb collaborators, we will continue to work to dissect pathways and delineate molecular mechanisms governing diverse CRISPR-Cas systems. Furthermore, we will gain key insight on ACR-mediated mechanisms used by viruses to thwart CRISPR immunity. Innovative CRISPR-based tools and technologies are providing transformative advances in basic and medical research. The knowledge gained by our research will contribute directly to ongoing efforts aimed at exploiting diverse CRISPR-Cas systems as powerful research tools (e.g. for genome editing and controlled gene expression, to selectively detect and target human viral and bacterial pathogens, and for limiting the spread of antibiotic resistance). Moreover, understanding the detailed modes of action of diverse ACRs is necessary for establishing ACRs as important new research tools for selective and safe utilization of different CRISPR-based technologies.