Abstract
Targeted genetic engineering of human pluripotent cells is a prerequisite for exploiting their full potential. Such genetic manipulations can be achieved using site-specific nucleases. Here we engineered transcription activator-like effector nucleases (TALENs) for five distinct genomic loci. At all loci tested we obtained human embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC) clones carrying transgenic cassettes solely at the TALEN-specified location. Our data suggest that TALENs employing the specific architectures described here mediate site-specific genome modification in human pluripotent cells with similar efficiency and precision as do zinc-finger nucleases (ZFNs).
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Base Sequence
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Embryonic Stem Cells / physiology*
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Endonucleases / genetics
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Endonucleases / metabolism*
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Gene Targeting / methods*
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Genetic Engineering / methods*
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Homeodomain Proteins / genetics
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Humans
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Induced Pluripotent Stem Cells / physiology*
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Molecular Sequence Data
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Myosin-Light-Chain Phosphatase / genetics
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Octamer Transcription Factor-3 / genetics
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Transcription Factors / genetics
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Transcription Factors / metabolism*
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Zinc Fingers
Substances
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Homeodomain Proteins
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Octamer Transcription Factor-3
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POU5F1 protein, human
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Transcription Factors
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homeobox protein PITX3
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Endonucleases
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Myosin-Light-Chain Phosphatase