Fig. 2

a Different approaches to pooled CRISPR-based screening. sgRNA libraries are introduced into a population of cells. Next, subpopulations are selected based on their proliferation/survival or based on physical separation by fluorescence-activated cell sorting (FACS) or magnetic-activated cell sorting (MACS) based on phenotypes of interest, and sgRNA frequencies in different populations are compared by targeted next-generation sequencing. Alternatively, phenotypes of interest are evaluated by microscopy, followed by photoactivation of a fluorescent protein in cells of interest to enable FACS sorting, or by in situ sequencing to identify sgRNAs. Lastly, pooled screens can be read out by single-cell RNA sequencing to identify both the sgRNA expressed in an individual cell and the transcriptomic consequences of gene perturbation. b Arrayed CRISPR-based screening. Here, different sgRNAs are individually introduced into cells, enabling additional readouts, including high-content imaging and monitoring of cell non-autonomous phenotypes