NK cells were edited with CRISPR as previously described (Riggan et al., 2020 (link)). Briefly, splenocytes were cultured with 50 ng/ml IL-15 overnight and electroporated with Cas9 complexed with NK1.1 (5′-GAGGAAGGTCAAGCTGACTG-3′; Riggan et al., 2020 (link)) or Ska2 (5′-GGTCGATAAGCTGGAACTGA-3′) gRNA selected using CRISPick (Doench et al., 2016 (link); all from Synthego) using Neon transfection (Thermo Fisher Scientific). Electroporated splenocytes were cultured for 4 d in the presence of 50 ng/ml IL-15 and NK cells were subsequently analyzed by flow cytometry.
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