GSM9251252: 7 genes KD in K562 (CRISPR); Homo sapiens; RNA-Seq
After pooled gRNA lentivirus transduction, cells were collected in PBS + 5% FBS and FACS sorting for mCherry (dCas9-KARB) and TagBFP (sgRNA) double-positive population. Sorted cells were cultured with fresh medium maintaining at a density of 100k cells/ml. Seven days after pooled sgRNA transduction (day 7), cells were sorted again for viability and double positive. The harvested cells were sent for sequencing immediately. Single cells were collected for the construction of 5' single-cell gene expression (GEX) libraries and CRISPR guide RNA (gRNA) capture libraries using the Chromium X platform (10x Genomics, PN-1000287, PN-1000451, PN-1000263, PN-1000215). The libraries were prepared with a target recovery of approximately 10,000 cells. Both GEX and CRISPR libraries were sequenced on an Illumina NovaSeq 6000 system using a 100-cycle kit.