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Developmental Biology

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JoVE Journal Developmental Biology
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

Article DOI: 10.3791/52504-v • 07:28 min • February 1st, 2015
February 1st, 2015 •
1, 1, 2, 1
1National Institute of Arthritis, Musculoskeletal and Skin Diseases, National Institutes of Health, 2Q Therapeutics

Chapitres

résumé
Automatic Translation

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

Transfection Selection Colony-picking Induced Pluripotent Stem Cells TALEN-targeted GFP Gene AAVS1 Safe Harbor Transgene Addition Gene Expression Viral Vector Gene Silencing Insertional Mutagenesis Open-source TALENs Gene-trap-like Donor Delivery Efficiency IPSC Preparation Nuclear Delivery Efficiency Gentle-cell Dissociation Reagent Optimized Program Gene Editing Vectors AAVS1 Locus PPP1R12C Gene Splicing Acceptor (SA) Puromycin Resistant Gene (PAC) Targeted IPSCs Random Integration-only Untransfected Cells
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