GSM7897841: Jurkat HIV, stimulated cells, GFP+; Homo sapiens; RNA-Seq
Transduction was carried out on 850,000 cells at an MOI of 0.3 using 8ug/ml polybrene in 2 ml of RPMI containing 10% FBS and 1% penicillin-streptomycin. The media was replaced 24 hours later with fresh media without polybrene. Two days after transduction, the cells were selected for using 1.5 ug/ml puromycin for 5 days. After selection, the cell were split evenly into three groups. One third of the cells were kept in culture with no drug added, while two thirds of the cells were stimulated with PMA/I. After 16 hours, the stimulated cells were sorted into GFP+ and GFP- populations. We designed Perturb-seq library that targets 10 protein-coding genes, which are either (1) known factors in HIV-1 virus expression and T cell activation (e.g., BIRC2), or (2) top hits from genome-scale CRISPR screens that we previously performed (e.g., BRD4).