RNA and ATAC modalities of single-cell dual-modal CRISPR screen of Homo sapiens: A375 melanoma Vemurafenib resistance
A375-dCas9-BFP-KRAB cells were transduced with a sgRNA library at a low MOI and BFP+ GFP+ double positive cells were sorted out. Cells were cultured for additional 6 days with either DMSO or 1uM Vemurafenib treatment. Fixed cells underwent Tn5 tagmentation in bulk, reverse transcription in bulk, two rounds of indexing ligation, and finally indexing PCR. SgRNA libraries were further separated and enriched by another round of PCR. A step-by-step experimental protocol of PerturbFate library preparation is included in the supplementary material of the publication. Both ATAC and RNA parts of the dual omics libraries share the cell barcodes, so they are included in the same raw fastq and replies on computational demultiplexing of modality-specific reads.