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Generación de células NK humano primario y ampliado de Knock-out utilizando ribonucleoproteínas Cas9
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면역학 및 감염병학
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JoVE 신문 면역학 및 감염병학
Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
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Generación de células NK humano primario y ampliado de Knock-out utilizando ribonucleoproteínas Cas9

12,174 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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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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Aquí, presentamos un protocolo para modificar genéticamente las células primarias o ampliadas humano naturales del asesino (NK) usando Cas9 ribonucleoproteínas (Cas9/RNPs). Mediante este protocolo, hemos generado las células NK humanas deficientes para el receptor 2 del factor de crecimiento-b de transformación (TGFBR2) y la hipoxantina fosforribosiltransferasa 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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