Targeted genome editing in pluripotent stem cells using zinc-finger nucleases

Xenopatients 2.0: Reprogramming the epigenetic landscapes of patient-derived cancer genomes

“…This method would test the sufficiency of the mutation for disease, the necessity of the mutation for the disease behavior, and/or the response to molecularly targeted drugs, and correct key disease mutations in a cancer patientspecific iPS cell line. Currently existing genomic-editing tools in human pluripotent stem cells, e.g., zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced shored palindromic repeats (CRISPRs)/CRISPR-associated (Cas) systems, [67][68][69][70][71] could perform proofof-principle exercises to unambiguously elucidate the necessity and sufficiency of given mutations for given cancer disease phenotypes. This approach might be extremely valuable to assess the importance of genetic modifiers on cancer disease penetrance, i.e., whether a cancer mutation evokes a diseases phenotype in some cell lines but not in others.…”

Human iPSC Models: A Platform for Investigating Neurodevelopmental Diseases

“…Hence, ZFN pairs require extensive in vitro testing making them an expensive choice for genome editing. ZFN genome editing is more fully reviewed in [118,119].…”

Generation and Genetic Engineering of Human Induced Pluripotent Stem Cells Using Designed Zinc Finger Nucleases

“…Targeted CFTR gene addition to the CCR5 safe harbor locus of the human genome Genetic engineering of hiPSCs using designed ZFNs or TALENs have been reported previously in literature employing hiPSCs generated by random integrations of pluripotency genes [21][22][23][24][25] using retroviral methods, rather than site-specific integration. ZFN-mediated approach, however, generates precisely targeted genetically well-defined hiPSCs, circumventing the problems associated with random integrations.…”

“…ZFN-mediated approach, however, generates precisely targeted genetically well-defined hiPSCs, circumventing the problems associated with random integrations. Targeting of selectable marker transgenes (like GFP or puromycin) to the IL2RG, PPP1R12C, AAVS1, and CCR5 loci have been described previously elsewhere [21,24,25].…”

“…Published reports from several labs have now firmly established ZFN-mediated gene targeting as a powerful research tool for site-specific and precise modification of the human genome. Site-specific modification of the human genome using designed ZFNs [and more recently using transcription activator like effector nucleases (TALENs)], has been successfully demonstrated in a variety of human cells and other cell types [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24]. High rate of endogenous gene modification efficiencies ( > 10%) have been achieved using ZFN-driven approach [8].…”