scCRISPR-screen in human monocyte derived macropahges

To determine whether the hits derived from a genome-wide murine BMDM screen regulated human macrophage phenorypes, we established a framework that coupled genetic perturbation with phenotypic analysis at single-cell resolution in primary human monocyte-derived macrophages (MDM scCRISPR-seq). MDM scCRISPR-seq combines highly efficient electroporation-based CRISPR gene editing with a perturbation-specific barcoding strategy using cell hashing antibodies, allowing for a pooled sequencing format of 84 conditions per donor. Our sgRNA library consisted of 4 sgRNAs per gene, targeting previsously identified hits in murine BMDMs along with negative controls including non-targeting controls (NTC) as well as positive controls of the interferon gamma signaling pathway (IFNGR1, SOCS1). CRISPR perturbed cells derived from two healthy donors were subjected to IFNγ stimulation or left untreated followed by cell hashing, pooling and scRNAseq analysis ***Raw data will be deposited at EGA***