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Immunology and Infection

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Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
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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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Knock-out Primary NK Cells Expanded NK Cells Cas9 Ribonucleoproteins Gene Function Effector Cell Function High Success Rate DNA-dependent Approaches Clinically Relevant Genes NK Cell Therapy Immunotherapy Solid Tumors Brain Tumors TGF-beta Expression Transduction RPMI Medium IL2 Supplementation T75 Flask IL21-expressing K562 Feeder Cells Electroporation
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