title: sgRNA amplicon of primary human NK cells transduced with CRISPRko kinome library
design: The kinome CRISPR knockout screen was performed in primary NK cells using the SLICE (sgRNA lentiviral infection with Cas9 protein electroporation) approach developed for primary T cells (Shifrut et al., 2018). Human Kinome CRISPR Knockout Library was from Addgene (catalog #1000000082) and prepared as described (Doench et al., 2016). The kinome CRISPR sgRNA library was packaged into BaEV-LV in the ASCT2 KO cells using our standard protocol. The viral titer was determined in HEK293T cells by puromycin selection as described above. About 1  107 of the feeder-expanded NK cells were transduced with the BaEV-LV at MOI 0.3. Recombinant Cas9 protein was prepared as detailed in (Lin et al., 2022) At 24 hours after transduction, Cas9 was electroporated to the NK cells using Lonza 4D nucleofection system using pulse code CM137 in 100-L electroporation cuvettes. The reaction contained 400 pmol of Cas9 protein and 1  107 of the transduced NK cells in 100 L of P3 solution (Lonza).