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Transfection, selezione, e Colonia-picking di cellule staminali umane pluripotenti indotte TALEN-mirati con Gene GFP nel AAVS1 Safe Harbor
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Chapitres
- 00:05Titre
- 01:31Inducible Pluripotent Stem (iPS) Cell Transfection
- 03:50Colony Picking and Targeted iPSC Expansion
- 05:37Results: Representative Gene-targeted iPSCs
- 07:01Conclusion
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
