Seamless genome editing in human pluripotent stem cells using custom endonuclease–based gene targeting and the piggyBac transposon

@article{Yusa2013SeamlessGE,
  title={Seamless genome editing in human pluripotent stem cells using custom endonuclease–based gene targeting and the piggyBac transposon},
  author={Kosuke Yusa},
  journal={Nature Protocols},
  year={2013},
  volume={8},
  pages={2061-2078}
}
I report here a detailed protocol for seamless genome editing using the piggyBac transposon in human pluripotent stem cells (hPSCs). Recent advances in custom endonucleases have enabled us to routinely perform genome editing in hPSCs. Conventional approaches use the Cre/loxP system that leaves behind residual sequences in the targeted genome. I used the piggyBac transposon to seamlessly remove a drug selection cassette and demonstrated safe genetic correction of a mutation causing α-1… CONTINUE READING
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