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

CRISPR-Cas是一种广泛存在的微生物免疫系统，可以对抗寄生的外源DNA。他们还催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基因修饰的能力。本项目将研究CRISPR- cas3 (Type I CRISPR- cas system)，这是自然界中发现的数量最多、系统发育最多样化的CRISPR分支。通过利用Cas9无法获得的I型CRISPR的独特功能，该项目具有开发具有前所未有的靶向灵活性的新技术和应用的潜力，这将显著推进人类基因组研究。此外，对原核生物遗传干扰途径的多样性将产生新的生物学见解。随着CRISPR技术继续改变农业、医学和生物技术部门，使公众广泛了解其科学和社会影响是一项重要的任务。该项目包括通过以crispr为主题的博物馆展览和高中教育计划进行公众宣传，以使下一代多样化的年轻思想家能够从事科学和技术事业。I型CRISPR的酶促机制具有Cas9无法获得的许多分子特性，但在生物技术中尚未开发。本研究的总体研究目标是促进对不同类型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)通过建立全新类型的I型效应物并阐明它们介导细菌防御的机制，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多亚基CRISPR-Cas系统的多样性、功能和生物技术潜力的理解。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提供支持。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.