Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 RibonucleoproteinsMeisam Naeimi Kararoudi 1, Hamid Dolatshad 2, Prashant Trikha 1, Syed-Rehan A. Hussain 3, Ezgi Elmas 1, Jennifer A. Foltz 1, Jena E. Moseman 1, Aarohi Thakkar 1, Robin J. Nakkula 1, Margaret Lamb 1, Nitin Chakravarti 1, K. John McLaughlin 3, Dean A. Lee 1
1Center for Childhood Cancer and Blood Disease, Nationwide Children's Hospital, 2Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, 3Center for Clinical and Translational Research, Nationwide Children's Hospital Research Institute
Here, we present a protocol to genetically modify primary or expanded human natural killer (NK) cells using Cas9 Ribonucleoproteins (Cas9/RNPs). By using this protocol, we generated human NK cells deficient for transforming growth factor–b receptor 2 (TGFBR2) and hypoxanthine phosphoribosyltransferase 1 (HPRT1).