Extended Data Fig. 1: ReLiC library design and validation.
From: Decoding post-transcriptional regulatory networks by RNA-linked CRISPR screening in human cells
a. Validation of Cas9 activity in U2OS. sgEYFP and sgCTRL are single guide RNAs targeting EYFP or a non-targeting control, respectively. Each histogram represents fluorescence of 10,000 cells as measured by flow cytometry. ‘Days post Cas9’ refers to days after addition of doxycycline to induce Cas9 expression. b.Comparison of integration into 293T and U2OS landing pads. BFP and mCherry fluorescence were measured for 10,000 cells, depicted as individual points. Proportion of cells that are mCherry+ and BFP- (orange points) is indicated. No cells in either parental control are mCherry+ and BFP-. c. Depiction of cloning scheme for ReLiC library and reporters. d. Distribution of sgRNA-linked barcode counts in mRNA and genomic DNA. e. Number of unique barcodes linked to each sgRNA in ReLiC library. f. Correlation between distinct barcode sets in ReLiC fitness screens. Each point represents a unique sgRNA pair from the ReLiC RBP library. For each sgRNA pair, individual linked barcodes were randomly partitioned into two sets of equal size (or to within a barcode for odd number of detected barcodes). r refers to Pearson correlation coefficient between the barcode sets.
