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Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells
JoVE Journal
Biology
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JoVE Journal Biology
Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells
DOI:

12:13 min

August 23, 2014

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Chapters

  • 00:05Title
  • 02:24Cell Preparation
  • 04:40Electroporation of hESCs
  • 07:34Picking Transgenic hESC Clones
  • 09:24Screening and Expansion of Positive Clones
  • 10:22Representative Screening Results
  • 11:41Conclusion

Summary

Automatic Translation

Reporter cell lines offer a means to visualize, track and isolate cells of interest from heterogeneous populations. However, gene-targeting using conventional homologous recombination in human embryonic stem cells is extremely inefficient. Herein, we describe targeting CNS midbrain specific transcription factor PITX3 locus with EGFP using zinc-finger nuclease enhanced homologous recombination.

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