Joint AstraZeneca-Cancer Research Horizons Functional Genomics Centre's CRISPRn library benchmark screens: gRNA counts and associated metadata

Genome-wide CRISPR sgRNA libraries have emerged as transformative tools to systematically probe gene function. While these libraries have been iterated over time to be more efficient, their large size limits their use in some applications. Here, we benchmarked publicly available genome-wide single-targeting sgRNA libraries and evaluated dual targeting as a strategy for pooled CRISPR loss-of-function screens. We leveraged this data to design two minimal genome-wide human CRISPR-Cas9 libraries that are 50% smaller than other libraries and that preserve specificity and sensitivity, thus enabling broader deployment at scale.全基因组CRISPR单引导RNA（single-guide RNA, sgRNA）文库已成为系统性探究基因功能的变革性工具。尽管此类文库历经多轮迭代以提升筛选效能，但其庞大的文库规模仍限制了部分应用场景的使用。本研究对公开可用的全基因组单靶向sgRNA文库进行了性能基准测试，并评估了双靶向策略在池化CRISPR功能丧失筛选中的应用效果。基于上述实验数据，本研究设计了两款极简型全基因组人类CRISPR-Cas9文库，其文库规模较现有同类文库缩减50%，同时保留了原有的特异性与检测灵敏度，从而可实现更大范围的规模化推广应用。