Single-Cell CRISPR Screening CapitalBiotech

Traditional CRISPR screening typically begins with the construction of a pooled sgRNA library containing hundreds to thousands of sgRNAs. The library is then introduced into cells stably expressing Cas9 to enable large-scale gene knockout screening. Cells are subjected to phenotype screening and bulk sequencing to obtain information on sgRNA changes during the culture process, so as to know the impact of the knocked-out gene on the cells. Conventional pooled CRISPR screening is well suited to phenotypes such as cell survival or proliferation, but provides limited resolution for studying molecular phenotypic changes at the single-cell level. To address this limitation, the 10x Genomics single-cell CRISPR screening platform combines pooled CRISPR perturbation with high-throughput single-cell sequencing, enabling simultaneous measurement of gene expression profiles and sgRNA identities in individual cells. This approach links genetic perturbations to molecular phenotypes at single-cell resolution.