Department of Pharmacology & Toxicology, University of Utah, Salt Lake City, UT, USA.
Methods Mol Biol. 2024;2707:319-334. doi: 10.1007/978-1-0716-3401-1_21.
CRISPR-Cas9 is a powerful tool to interrogate gene function in a targeted and systematic manner. Although the technology has been scaled up for large-scale genetic screens in cell culture, similar scale screens in vivo have been extremely challenging due to the cost, labor, and time required to generate and keep track of thousands of mutant animals. We reported the development of Multiplexed Intermixed CRISPR Droplets (MIC-Drop), a platform that makes large-scale reverse genetic screens possible in zebrafish. In this chapter, we provide a detailed protocol to conduct large-scale genetic screens using this novel platform.
CRISPR-Cas9 是一种强大的工具，可以有针对性和系统性地研究基因功能。尽管该技术已经在细胞培养中进行了大规模的遗传筛选，但由于生成和跟踪数千种突变动物所需的成本、劳动力和时间，在体内进行类似规模的筛选极具挑战性。我们报告了多重混合 CRISPR 液滴（MIC-Drop）平台的开发，该平台使在斑马鱼中进行大规模反向遗传筛选成为可能。在本章中，我们提供了使用该新型平台进行大规模遗传筛选的详细方案。
