Nuclease-mediated Double Strand Break (DSB) Enhancement of Small Fragment Homologous Recombination (SFHR) Gene Modification in Human Induced Pluripotent Stem Cells (hiPSCs)
| Autor: | Dieter C. Gruenert, R. Geoffrey Sargent, Shingo Suzuki |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Transcription activator-like effector nuclease
Cystic Fibrosis DNA Repair DNA repair Genetic enhancement Induced Pluripotent Stem Cells Cystic Fibrosis Transmembrane Conductance Regulator Biology Endonucleases Molecular biology Article chemistry.chemical_compound chemistry Transcription (biology) Gene Targeting Humans DNA Breaks Double-Stranded Induced pluripotent stem cell Homologous recombination Homologous Recombination Gene DNA |
| Zdroj: | Methods in Molecular Biology ISBN: 9781627037600 |
| Popis: | Recent developments in methods to specifically modify genomic DNA using sequence-specific endonucleases and donor DNA have opened the door to a new therapeutic paradigm for cell and gene therapy of inherited diseases. Sequence-specific endonucleases, in particular transcription activator-like (TAL) effector nucleases (TALENs), have been coupled with polynucleotide small/short DNA fragments (SDFs) to correct the most common mutation in the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene, a 3-base-pair deletion at codon 508 (delF508), in induced pluripotent stem (iPS) cells. The studies presented here describe the generation of candidate TALENs and their co-transfection with wild-type (wt) CFTR-SDFs into CF-iPS cells homozygous for the delF508 mutation. Using an allele-specific PCR (AS-PCR)-based cyclic enrichment protocol, clonal populations of corrected CF-iPS cells were isolated and expanded. |
| Databáze: | OpenAIRE |
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