CAREER: Expanding and Leveraging the Natural Diversity of Type I CRISPR for Genome Engineering

CRISPR-Cas是针对寄生性外源DNA的广泛的微生物免疫系统。他们还产生了获得诺贝尔奖的Cas9基因编辑工具，这些工具彻底改变了在植物和动物中安装可编程遗传修饰的能力。该项目将研究CRISPR-Cas 3（I型CRISPR-Cas系统），这是自然界中发现的最丰富和最具遗传多样性的CRISPR分支。通过利用Cas9无法获得的I型CRISPR的独特功能，该项目有可能开发具有前所未有的靶向灵活性的新技术和应用，这将大大推进人类基因组研究。此外，新的生物学见解将产生关于原核遗传干扰途径的多样性。随着CRISPR技术不断改变农业、医药和生物技术领域，让公众了解科学和社会影响是一项重要的任务。该项目包括通过CRISPR主题的博物馆展览和高中教育项目进行公众宣传，使下一代多样化的年轻思想家能够追求科学和技术事业。I型CRISPR的酶机制具有Cas9无法获得的许多分子特性，但在生物技术方面尚未开发。本研究的总体研究目标是推进对不同I型CRISPR-Cas系统的基本理解，并扩大其在人类基因组工程中的应用。该项目将追求两个研究目标，包括（1）将现有的I型CRISPR变体开发成新的基因编辑平台，以扩大当前CRISPR工具包的靶向范围和应用;（2）通过建立全新的I型效应子类别并阐明它们介导细菌防御的机制来扩大I型CRISPR的自然多样性。一个多方面的方法混合人类细胞培养，高通量测序和细菌遗传学将被采用。这项工作有可能改变我们对生物多样性，功能，和多亚基CRISPR的生物技术潜力-Cas系统。该项目由生物科学理事会分子和细胞生物科学部的遗传机制计划和系统与合成生物学计划支持。该奖项反映了NSF的法定使命，并通过使用基金会的学术价值和更广泛的影响评审标准。

CRISPR-Cas are widespread microbial immune systems against parasitic foreign DNA. They also gave rise to the Nobel Prize-winning Cas9 gene-editing tools that have revolutionized the ability to install programmable genetic modifications in plants and animals. This project will investigate CRISPR-Cas3 (Type I CRISPR-Cas system), which is the most abundant and phylogenetically diverse CRISPR branch found in nature. By exploiting unique features of Type I CRISPR unavailable to Cas9, this project has the potential to develop new technologies and applications with unprecedented targeting flexibility, which will significantly advance human genome research. Furthermore, new biological insights will be generated about the diversity of prokaryotic genetic interference pathways. As CRISPR technologies continue to transform agriculture, medicine, and biotechnology sectors, enabling broad public understanding of the scientific and societal impacts is an important undertaking. This project includes public outreach through CRISPR-themed museum exhibitions and high school educational programs, to empower the next generation of diverse young thinkers to pursue careers in science and technology.The enzymatic machinery of Type I CRISPR has many molecular properties unavailable to Cas9 but is under-developed in biotechnology. The overall research goal of this study is to advance the fundamental understanding of divergent Type I CRISPR-Cas systems and broaden their utilities in human genome engineering. This project will pursue two research objectives, including (1) the development of existing Type I CRISPR variants into novel gene-editing platforms to expand the targeting scope and applications of the current CRISPR toolkit; and (2) the expansion of Type I CRISPR’s natural diversity by establishing brand-new classes of Type I effectors and elucidating the mechanisms whereby they mediate bacterial defense. A multifaceted approach blending human cell culture, high-throughput sequencing, and bacterial genetics will be employed. This work has the potential to transform our understanding of the diversity, function, and biotechnological potential of multi-subunit CRISPR-Cas systems.This project is supported by the Genetic Mechanisms program and the Systems and Synthetic Biology program of the Division of Molecular and Cellular Biosciences in the Directorate for Biological Sciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.