GSM8817950: iSAM hESCs after CRISPRa screen interrogation to discover CCRs using high MOI gRNA library (gRNA library); Homo sapiens; RNA-Seq
mCherry+ hESCs transduced with the 4 different library conditions were FACS-sorted and 2 cellular mixes were prepared as it follows: 1.) 500000 cells at 97:3 ratio = low MOI candidate CCR-library transduced hESCs : low MOI negative control transduced hESCs. 2.) 100000 cells at 97:3 ratio = high MOI candidate CCR-library transduced hESCs : high MOI negative control transduced hESCs. Cellular suspensions were loaded on a Chromium Controller targeting 30000 cells per reaction. Seven reactions for the low MOI condition and one reaction for the high MOI condition were loaded. Single-cell 3′ transcriptome libraries were generated following the manufacturer's instructions (10x Genomics Chromium Single Cell 3′ Reagent Kit v3.1-Dual Index User Guide). gRNA libraries were generated by amplifying the fragment that contains the gRNA sequence from the MS2-CROP construct (see oligos in table S9), and a second PCR to add index adaptors from the Chromium Single Cell 3′ Reagent Kit v3.1-Dual Index.