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Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus
JoVE Journal
Medicine
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JoVE Journal Medicine
Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus

Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus

DOI:
10.3791/54718-v

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11:36 min

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November 20, 2016

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, , , , , , , , , , ,
1Stem Cell Institute,Katholieke Universiteit Leuven

Chapters

  • 00:05Title
  • 00:58Preparation of FRT-containing hPSC Line for Transfection
  • 04:44Transfection by Nucleofection
  • 06:28Positive and Negative Selection of Cells Undergoing RMCE
  • 07:28Expansion and Characterization of RMCE Lines
  • 08:31Results: RMCE in the AAVS1 of hPSC
  • 10:51Conclusion

Summary

Automatic Translation

Automatic Translation

Here we report a rapid and efficient gene editing method based on RMCE in the AAVS1 locus of human Pluripotent Stem Cells (hPSCs) that improves upon previously described systems. Using this technique, isogenic lines can be rapidly and reliably generated for proper comparative studies, facilitating transgenesis-mediated research with hPSCs.

Tags

Keywords: RecombinantHuman Pluripotent Stem CellsAAVS1 LocusRecombinase-mediated Cassette ExchangeRMCETransgenesisStem Cell BiologyOverexpressionKnockdownRandom IntegrationKaryotypesIMEFsROCK InhibitorNucleofectionTrypsinAccutaseFeeder-free

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