CROP-seq + PIP-seq

We used PIP-seq with a CROP-seq library to evaluate the feability of PIP-seq for large pooled CRISPR screens Overall design: K562 CRISPRi cells were cultured in RPMI-1640 (Gibco #11875093) with 10% FBS (Thermo Fisher Scientific, #10438026) and 1% penicillin/streptomycin (Thermo Fisher Scientific, #15140148) in an incubator at 37°C with 5% CO2. K562 CRISPRi cells were transduced with a lentivirus library containing 138 sgRNA (Jost et al 2020) at a multiplicity of infection of 0.1. Lentivirus infected cells (BFP+) were sorted to high purity using a BD FACS Aria III (100 µMnozzle) and processed according to the PIP-seq scRNA-seq workflow.