Robertson Nicholas R, Lee Sangcheon, Tafrishi Aida, Wheeldon Ian
Bioengineering, University of California, Riverside, Riverside, CA 92521, United States.
Chemical and Environmental Engineering, University of California, Riverside, Riverside, CA 92521, United States.
FEMS Yeast Res. 2025 Jan 30;25. doi: 10.1093/femsyr/foaf013.
Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas genome-wide screens are powerful tools for unraveling genotype-phenotype relationships, enabling precise manipulation of genes to study and engineer industrially useful traits. Traditional genetic methods, such as random mutagenesis or RNA interference, often lack the specificity and scalability required for large-scale functional genomic screens. CRISPR systems overcome these limitations by offering precision gene targeting and manipulation, allowing for high-throughput investigations into gene function and interactions. Recent work has shown that CRISPR genome editing is widely adaptable to several yeast species, many of which have natural traits suited for industrial biotechnology. In this review, we discuss recent advances in yeast functional genomics, emphasizing advancements made with CRISPR tools. We discuss how the development and optimization of CRISPR genome-wide screens have enabled a host-first approach to metabolic engineering, which takes advantage of the natural traits of nonconventional yeast-fast growth rates, high stress tolerance, and novel metabolism-to create new production hosts. Lastly, we discuss future directions, including automation and biosensor-driven screens, to enhance high-throughput CRISPR-enabled yeast engineering.
成簇规律间隔短回文重复序列（CRISPR）-Cas全基因组筛选是揭示基因型-表型关系的强大工具，能够精确操纵基因以研究和设计具有工业用途的性状。传统的遗传方法，如随机诱变或RNA干扰，往往缺乏大规模功能基因组筛选所需的特异性和可扩展性。CRISPR系统通过提供精确的基因靶向和操纵克服了这些限制，从而能够对基因功能和相互作用进行高通量研究。最近的研究表明，CRISPR基因组编辑广泛适用于多种酵母物种，其中许多酵母具有适合工业生物技术的天然特性。在本综述中，我们讨论了酵母功能基因组学的最新进展，重点强调了CRISPR工具所取得的进展。我们讨论了CRISPR全基因组筛选的开发和优化如何实现了一种宿主优先的代谢工程方法，该方法利用非常规酵母的天然特性——快速生长速率、高胁迫耐受性和新型代谢——来创建新的生产宿主。最后，我们讨论了未来的方向，包括自动化和生物传感器驱动的筛选，以加强高通量CRISPR介导的酵母工程。
