Fig. 1: A genome-wide pooled CRISPR screen identifies novel RNA-binding proteins which regulate TfR1 expression.

a, b Workflow (a) and cell sorting strategy (b) of the genome-wide CRISPR screen for TfR1 regulators. c Gene ontology analysis of the TfR1 CRISPR screen hits. Statistical analysis was performed using Immuno-Navigator⁸³. d Volcano plot of TfR1 phenotype (casTLE Effect, the most likely effect size as determined by casTLE) and the confidence in that effect size (casTLE Score) from the CRISPR screen. The screen was performed in duplicate. e–g Validation of the screen hits. Two gRNAs per gene were chosen for retests. K562-Cas9 cells were transduced with lentiviral vectors encoding individual gRNAs together with mCherry. Then cells were stained with anti-TfR1 antibody and analyzed using flow cytometry (e). Change in TfR1 expression levels between untransduced (mCherry−, gray) and transduced (mCherry+, red) were calculated and summarized (f, g). Each symbol represents the results of individual gRNAs. Data were pooled from two independent experiments. h, i Effects of wild-type or catalytic-dead mutant METTL16 (PP185/186AA, F187G) on surface TfR1 expression in the presence of DFO. Similar expression levels of wild-type or mutant METTL16 protein were confirmed (h). Surface TfR1 expression levels of cells expressing indicated gRNAs and proteins were examined by flow cytometry (i). Similar results were obtained in two independent experiments. Source data are provided as a Source Data file.