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

English

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12,183 Views

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07:20 min

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June 14, 2018

DOI:

10.3791/58237-v

DOI:
10.3791/58237-v

•

07:20 min
June 14, 2018

•

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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

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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Naeimi Kararoudi, M., Dolatshad, H., Trikha, P., Hussain, S. A., Elmas, E., Foltz, J. A., Moseman, J. E., Thakkar, A., Nakkula, R. J., Lamb, M., Chakravarti, N., McLaughlin, K. J., Lee, D. A. Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins. J. Vis. Exp. (136), e58237, doi:10.3791/58237 (2018).

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