Shortly, peripheral blood mononuclear cells (PBMCs) were thawed and activated using soluble anti-CD2/3/28 (Stem Cell Technology). At day 3 post-activation, 1x10^6 cells were nucleofected (4D-Nucleofector, Lonza; kit P3, program EO-115) with 20 pmol of SpCas9 protein (PNA Bio) complexed with 100 pmol of gRNA for 10 min at RT. T cells were cultured in 96 well plates (Corning, U bottom) up to 6 days before genotypic analyses. 1x10^6 pre-cultivated NK cells were nucleofected (4D-Nucleofector, Lonza; optimized NK cell nucelofection protocol recently described by Ullrich group 4) using the indicated amounts of Cas9:gRNA. NK cells were then expanded with IL-15 containing medium as recently described 3 up to two to three weeks before further analyses. MM1.S cells were cultured 2 days before genome editing. Afterward, 3 µg of SpCas9 protein (BioInc) was complexed with 100 pmol of gRNA for 10 min at RT, followed by nucleofection of 2x10^5 MM1.S cells (4D-Nucleofector, Lonza; kit SF, program CM-137).