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Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
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

DOI:
10.3791/58237-v

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07:20 min

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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

Capitoli

  • 00:04Titolo
  • 00:44Human Primary and Expanded Natural Killer (NK) Cell Transduction
  • 04:48Results: Representative Electroporation Efficiency
  • 06:13Conclusion

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).

Tags

Cas9 RibonucleoproteinsNK CellsGene KnockoutPrimary NK CellsExpanded NK CellsCRISPR RNATracrRNAElectroporationTransductionImmunotherapySolid TumorsBrain TumorsTGF-beta

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