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Geração de células NK humana primárias e expandida de Knock-out usando ribonucleoproteínas Cas9
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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

Geração de células NK humana primárias e expandida de Knock-out usando ribonucleoproteínas Cas9

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12,187 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

•

5 Views
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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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Aqui, apresentamos um protocolo para modificar geneticamente as células assassinas naturais humano primário ou expandido (NK) usando Cas9 ribonucleoproteínas (Cas9/RNPs). Usando este protocolo, geramos humanas células NK deficientes para transformar o receptor 2 do fator de crescimento – b (TGFBR2) e hipoxantina Fosforibosiltransferase 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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