title: GSM4388149: THP1_sgRNA_S11; Homo sapiens; OTHER
library_construction_protocol: THP-1 Cas9-expressing cells were transduced with pooled sgRNA lentivirus at a low MOI (~0.3). At day 12 post-transduction, cells were stained with DAPI (Sigma Aldrich #D9542-1MG) and sorted for live GFP positive cells. Subsequently, sorted cells were stained with human Fcblock for 10 mins. at room temperature and then incubated with TotalSeq-C antibodies, CD11B (BioLegends #301359) and CD16 (BioLegends #302065) for 30 mins. on ice. Samples were processed according to Chromium Next GEM Single Cell V(D)J v1.1 kit with modified steps referenced in ECCITE-seq protocol29. Libraries were pooled consisting of 85% cDNA, 10% guide-tag and 5% protein-tag and sequenced on Nova-Seq SP100 (Recipe 26 cycles read 1, 8 cycles index, and 91 cycles read 2). Oligonucleotide sequences of sgRNAs used for the library and primers for ECCITE-seq can be found in Supplementary Table 1
attributes.tag: GEO Accession
attributes.value: GSM4388149