GSM5008989: CRISPRa-hESC CRISPR gRNA capture library; Homo sapiens; RNA-Seq
Cells were lifted using Accutase (Gibco) and resuspended in FACS buffer (1X Dulbecco's  phosphate-buffer saline (DPBS; Lonza) supplemented with 0.1% (w/v) Bovine Serum Albumin (BSA; Sigma Aldrich) and 2.5mM EDTA (Invitrogen)). Cells were stained with Propidium Iodide (PI; 1:500; Sigma Aldrich) prior to sorting using the FACSAriaTM III cell sorter (BD Biosciences). Cells expressing gRNA of interest were defined as live/ PI-​ and GFP+. Control population was defined as live/ PI-​ ​. 30,000 cells per sample (10 samples) were sorted into FACS buffer for downstream processing for scRNA-seq. Single cell capture, RNA-seq library construction and sequencing were carried out using the Chromium Next GEM Single Cell 3' Reagent Kits v3.1 with Feature Barcoding technology (10X Genomics) single-cell RNA sequencing