Fig. 2: Design and functional validation of a new gRNA library tailored for CRISPR-knockout screens in human myoblasts.
a Design of the muscle-targeted CRISPR knockout library (MyoCRISPR-KOLib), comprising 20,496 gene-targeting gRNAs (~3 per gene) and 900 non-targeting control gRNAs. TPM: transcripts per million. b Cumulative mRNA expression levels of genes included in the library, shown as a percentage of total mRNA from all protein-coding genes, based on RNA-seq analysis of seven human myoblast lines. Diff.: differentiation. c Cumulative read count plot showing the uniform distribution of gRNA abundances in the MyoCRISPR-KOLib, as determined by deep sequencing. d Overview of the myoblast fitness screen. Each dot represents a single gRNA expression and corresponding gene perturbation. d’, Hypothetical results illustrate the potential impact of gene perturbation, interpreted by comparing gRNA abundance across samples from different time points. e Top three pathways enriched among the 419 genes (Supplementary Data 3) identified in the myoblast fitness screen. f gRNA quantification results for two representative hits. g Top three pathways enriched among the 47 negative regulators (Supplementary Data 4) identified in the myoblast fitness screen. h gRNA quantification results for two representative hits. Source data are provided as a Source data file.
