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Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor
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Biologia do Desenvolvimento
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JoVE Journal Biologia do Desenvolvimento
Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

DOI:
10.3791/52504-v

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07:28 min

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February 01, 2015

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, , ,
1National Institute of Arthritis, Musculoskeletal and Skin Diseases,National Institutes of Health, 2Q Therapeutics

Capítulos

  • 00:05Título
  • 01:31Inducible Pluripotent Stem (iPS) Cell Transfection
  • 03:50Colony Picking and Targeted iPSC Expansion
  • 05:37Results: Representative Gene-targeted iPSCs
  • 07:01Conclusion

Summary

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TALEN-mediated gene editing at the safe harbor AAVS1 locus enables high-efficiency transgene addition in human iPSCs. This protocol describes the procedures for preparing iPSCs for TALEN and donor vector delivery, transfecting iPSCs, and selecting and isolating iPSC clones to achieve targeted integration of a GFP gene to generate reporter lines.

Tags

Keywords: TransfectionSelectionColony-pickingHuman Induced Pluripotent Stem CellsTALENGFPAAVS1 Safe HarborTargeted Transgene AdditionGene EditingGene SilencingInsertional MutagenesisViral VectorGene-trap-like DonorSingle Cell DissociationNuclear Delivery EfficiencyPuromycin SelectionPPP1R12CSplicing AcceptorPACTargeted ClonesGenetic ScreeningFunctional ScreeningGFP Reporter IPSC Lines

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