GSM9864230: DLD1 CRISPRi sublib2, SAM24482910_4, MOI Low (~0.3), GEX; Homo sapiens; RNA-Seq
After 5 days of doxycycline and Shield-1 induction, cells from each MOI condition were harvested, labeled with MULTI-seq CMO hashtag barcodes, pooled, and sorted by FACS. Sorted cells were resuspended and loaded onto 10x Genomics Chromium GEM-X chips targeting ~50,000 cells per channel. Single-cell libraries were prepared using the 10x Genomics Chromium Single Cell 3' GEM-X v4 kit according to the manufacturer's instructions. Three libraries were constructed per 10x channel: gene expression (GEX), sgRNA capture, and MULTI-seq CMO (cell multiplexing oligo) hashing. For the scaled experiment (sub-library 2), 44 10x channels were used (Low MOI: 24, Medium MOI: 12, High MOI: 8). For the pilot experiment, 4 10x channels were used.