Extended Data Fig. 6: Technical properties of the ground truth perturbation experiment. | Nature Methods

Extended Data Fig. 6: Technical properties of the ground truth perturbation experiment.

From: Targeted Perturb-seq enables genome-scale genetic screens in single cells

Extended Data Fig. 6

a, Gene expression level in K562 cells of the various gRNA target genes used. b, Enhancer gRNAs were validated by pooled transduction of K562 dCas9-KRAB cells with all four enhancer-targeting guides, and the effect on target gene expression was quantified by qPCR. HBE1 was analyzed as target gene for the HS2 enhancer. n = 63 replicates. c, Histogram of the number of gRNAs identified per cell in the TAP-seq experiment of Fig. 2. d, The number of gRNAs observed per cell (see also in c) was fitted with a generative model of gRNA capture efficiency and multiplicity of infection4,20. Log-likelihood is plotted as a function of the parameters; the maximum likelihood estimate is marked by a cross. Data from n = 621,977 (TAP-seq), n = 67,994 (Perturb-Seq) or n = 637,971 cells (Perturb-seq + gRNA amp.) was used. e, Mean expression per gene for whole transcriptome 10X Genomics compared to TAP-seq, with perturbation target genes highlighted. n = 674 genes from panel 2 are shown. Two genes for which perturbation effects were detected with a lower efficiency in TAP-seq are highlighted in red.

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