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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
Medicina
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JoVE Journal Medicina
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

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11:36 min

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November 20, 2016

DOI:

10.3791/54718-v

DOI:
10.3791/54718-v

•

11:36 min
November 20, 2016

•

5 Views
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1Stem Cell Institute,Katholieke Universiteit Leuven

Summary

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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.

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

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Ordovás, L., Boon, R., Pistoni, M., Chen, Y., Sambathkumar, R., Helsen, N., Vanhove, J., Berckmans, P., Cai, Q., Vanuytsel, K., Raitano, S., Verfaillie, C. M. Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus. J. Vis. Exp. (117), e54718, doi:10.3791/54718 (2016).

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