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piggyBac Transposon System Modification of Primary Human T Cells
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Immunology and Infection
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JoVE Journal Immunology and Infection
piggyBac Transposon System Modification of Primary Human T Cells

piggyBac Transposon System Modification of Primary Human T Cells

DOI:
10.3791/4235-v

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10:02 min

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November 05, 2012

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, , , , , ,
1Program in Translational Biology and Molecular Medicine,Baylor College of Medicine , 2Department of Medicine, Division of Nephrology,Baylor College of Medicine , 3Department of Immunology and Pathology,Shinshu University School of Medicine, 4Center for Cell and Gene Therapy,Baylor College of Medicine , 5Department of Pediatrics,Baylor College of Medicine , 6Program in Cell and Molecular Biology,Baylor College of Medicine , 7Department of Molecular Virology and Microbiology,Baylor College of Medicine , 8Michael E. DeBakey VA Medical Center

Chapters

  • 00:05Title
  • 01:39Day 0: Isolation of PBMCs from Human Blood
  • 03:23Day 1: Coating Plates with anti-CD28 and anti-CD3 Antibody for Stimulating T Cells
  • 03:59Day 1: Nucleofection of Unstimulated T Cells
  • 05:54Day 2: Nonspecific Stimulation of T Cells
  • 07:28Day 7: Analysis of Gene Expression
  • 08:05Results: piggyBac-mediated Gene Modification of Human T Cells
  • 09:19Conclusion

Summary

Automatic Translation

Automatic Translation

We describe a method to genetically modify primary human T cells with a transgene using the non-viral piggyBac transposon system. T cells modified to using the piggyBac transposon system exhibit stable transgene expression.

Tags

PiggyBac Transposon SystemPrimary Human T CellsGene TransferPlasmidTransposase EnzymeInverted Repeat ElementsGene DeliveryAdoptive ImmunotherapyCancerClinical TrialNon-viral MethodologyGenetic ModificationReporter GeneInducible Suicide GeneGenomic Integration Sites

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