The paper details a comprehensive study employing genome-scale Perturb-seq screens—a technique combining genetic perturbations via CRISPR interference (CRISPRi) with single-cell RNA sequencing (scRNA-seq)—to create detailed genotype-phenotype maps in K562 and RPE1 cell lines. This methodology is shown to be highly effective for the unbiased assignment of gene function and for dissecting complex cellular phenotypes, such as RNA splicing, cell differentiation, and chromosomal instability (CIN). Key discoveries include the functional assignment of poorly characterized genes in processes like ribosome biogenesis and the identification of a novel submodule of the Integrator complex. Furthermore, the large-scale data allowed researchers to uncover previously unseen, stress-specific regulation of the mitochondrial genome.

References:

• Replogle J M, Saunders R A, Pogson A N, et al. Mapping information-rich genotype-phenotype landscapes with genome-scale Perturb-seq[J]. Cell, 2022, 185(14): 2559-2575. e28.