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Immunology and Infection

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JoVE Journal Immunology and Infection
Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
 

Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins

Article DOI: 10.3791/58237-v • 07:26 min • June 14th, 2018
June 14th, 2018 •
1, 2, 1, 3, 1, 1, 1, 1, 1, 1, 1, 3, 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

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Summary
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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).

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Keywords: Cas9 Ribonucleoproteins NK Cells Gene Knockout Primary NK Cells Expanded NK Cells CRISPR RNA TracrRNA Electroporation Transduction Immunotherapy Solid Tumors Brain Tumors TGF-beta
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