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Efficient Recombinase-Mediated Cassette Exchange in hPSCs to Study the Hepatocyte Lineage Reveals AAVS1 Locus-Mediated Transgene Inhibition

dc.contributor.authorLaura Ordovás
dc.contributor.authorRuben Boon
dc.contributor.authorMariaelena Pistoni
dc.contributor.authorYemiao Chen
dc.contributor.authorEsther Wolfs
dc.contributor.authorWenting Guo
dc.contributor.authorRangarajan Sambathkumar
dc.contributor.authorSylwia Bobis-Wozowicz
dc.contributor.authorNicky Helsen
dc.contributor.authorJolien Vanhove
dc.contributor.authorPieter Berckmans
dc.contributor.authorQing Cai
dc.contributor.authorKim Vanuytsel
dc.contributor.authorKristel Eggermont
dc.contributor.authorVeerle Vanslembrouck
dc.contributor.authorBéla Z. Schmidt
dc.contributor.authorSusanna Raitano
dc.contributor.authorLudo Van Den Bosch
dc.contributor.authorYaakov Nahmias
dc.contributor.authorToni Cathomen
dc.contributor.authorTom Struys
dc.contributor.authorCatherine M. Verfaillie
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherGroup of Morphology, Biomedical Research Institute, Hasselt University, Diepenbeek 3590, Belgium
dc.contributor.otherLeuven Research Institute for Neuroscience and Disease (LIND), Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherInstitute for Cell and Gene Therapy, University Medical Center Freiburg, Freiburg 79108, Germany
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherSwitch Laboratory, VIB, Leuven 3000, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.contributor.otherLeuven Research Institute for Neuroscience and Disease (LIND), Leuven 3000, Belgium
dc.contributor.otherDepartment of Cell and Developmental Biology, Hebrew University of Jerusalem, Jerusalem 91904, Israel
dc.contributor.otherInstitute for Cell and Gene Therapy, University Medical Center Freiburg, Freiburg 79108, Germany
dc.contributor.otherGroup of Morphology, Biomedical Research Institute, Hasselt University, Diepenbeek 3590, Belgium
dc.contributor.otherStem Cell Institute, KU Leuven, Leuven 3000, Belgium
dc.date.accessioned2015-11-12T05:00:39Z
dc.date.available2025-10-02T03:44:57Z
dc.date.issued01-11-2015
dc.identifier.issn-
dc.identifier.urihttp://www.sciencedirect.com/science/article/pii/S2213671115002660
dc.description.abstractTools for rapid and efficient transgenesis in “safe harbor” loci in an isogenic context remain important to exploit the possibilities of human pluripotent stem cells (hPSCs). We created hPSC master cell lines suitable for FLPe recombinase-mediated cassette exchange (RMCE) in the AAVS1 locus that allow generation of transgenic lines within 15 days with 100% efficiency and without random integrations. Using RMCE, we successfully incorporated several transgenes useful for lineage identification, cell toxicity studies, and gene overexpression to study the hepatocyte lineage. However, we observed unexpected and variable transgene expression inhibition in vitro, due to DNA methylation and other unknown mechanisms, both in undifferentiated hESC and differentiating hepatocytes. Therefore, the AAVS1 locus cannot be considered a universally safe harbor locus for reliable transgene expression in vitro, and using it for transgenesis in hPSC will require careful assessment of the function of individual transgenes.
dc.format-
dc.language.isoEN
dc.publisherElsevier
dc.relation.uri['https://wajhn.com/index.php/journal/editorial-policy', 'https://wajhn.com/index.php/journal/open-access-policy', 'https://wajhn.com/index.php/journal/index', 'https://wajhn.com/index.php/journal/aim-scope']
dc.rightsCC BY
dc.subject['health', 'medicine', 'public health', 'nutrition', 'health sciences', 'medical sciences', 'Nutritional diseases. Deficiency diseases', 'RC620-627']
dc.subject.lccMedicine (General)
dc.titleEfficient Recombinase-Mediated Cassette Exchange in hPSCs to Study the Hepatocyte Lineage Reveals AAVS1 Locus-Mediated Transgene Inhibition
dc.typeArticle
dc.description.pages918-931
dc.description.doi10.1016/j.stemcr.2015.09.004
dc.title.journalStem Cell Reports
dc.identifier.e-issn-
dc.identifier.oaioai:doaj.org/journal:104293e2640a470085393893e506221e
dc.journal.infoVolume 5, Issue 5


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