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An Inducible Expression System of the Calcium-Activated Potassium Channel 4 to Study the Differential Impact on Embryonic Stem Cells
Author(s) -
Stefan Liebau,
Michael Tischendorf,
Daniel Ansorge,
Leonhard Linta,
Marianne Stockmann,
Clair E. Weidgang,
Michelina Iacovino,
Tobias M. Boeckers,
Götz von Wichert,
Michael Kyba,
Alexander Kleger
Publication year - 2011
Publication title -
stem cells international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.205
H-Index - 64
eISSN - 1687-9678
pISSN - 1687-966X
DOI - 10.4061/2011/456815
Subject(s) - embryonic stem cell , stem cell , microbiology and biotechnology , doxycycline , potassium channel , cell , biology , calcium activated potassium channel , chemistry , gene , genetics , biophysics , antibiotics
Rationale. The family of calcium-activated potassium channels consists of four members with varying biological functions and conductances. Besides membrane potential modulation, SK channels have been found to be involved in cardiac pacemaker cell development from ES cells and morphological shaping of neural stem cells. Objective. Distinct SK channel subtype expression in ES cells might elucidate their precise impact during cardiac development. We chose SK channel subtype 4 as a potential candidate influencing embryonic stem cell differentiation. Methods. We generated a doxycycline inducible mouse ES cell line via targeted homologous recombination of a cassette expressing a bicistronic construct encoding SK4 and a fluorophore from the murine HPRT locus. Conclusion. We characterized the mouse ES cell line iSK4-AcGFP. The cassette is readily expressed under the control of doxycycline, and the overexpression of SK4 led to an increase in cardiac and pacemaker cell differentiation thereby serving as a unique tool to characterize the cell biological variances due to specific SK channel overexpression

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