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Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3
Paolo Cinelli1, Elisa A Casanova, Syndi Uhlig
1Institute of Laboratory Animal Science, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. paolo.cinelli@ltk.uzh.ch
BMC Developmental Biology
|May 27, 2008
Summary
STAT3 activation is crucial for maintaining embryonic stem cell pluripotency. Overexpressing STAT3-MER and specific downstream genes like Pem/Rhox5 and Pramel7 facilitates ES cell establishment and LIF-independent pluripotency.
Area of Science:
- Stem Cell Biology
- Molecular Biology
- Genetics
Background:
- Leukemia Inhibitory Factor (LIF) signaling is essential for maintaining the undifferentiated and pluripotent state of embryonic stem (ES) cells.
- Signal transducer and activator of transcription 3 (STAT3) is a key downstream target within the LIF signaling pathway.
- Investigating STAT3's role is critical for understanding ES cell pluripotency maintenance.
Purpose of the Study:
- To derive stable pluripotent ES cells from transgenic mice expressing a conditional STAT3-MER fusion protein.
- To identify novel genes involved in pluripotency maintenance through expression profiling of STAT3-activated ES cells.
- To validate the role of identified candidate genes in LIF-independent ES cell pluripotency.
Main Methods:
- Generation of transgenic FVB mice with a tamoxifen-dependent STAT3-MER fusion protein.
- Derivation of ES cells from STAT3-MER expressing blastocysts.
- Gene expression profiling (microarray or RNA-Seq) of STAT3-activated ES cells.
- Overexpression of candidate genes (Pem/Rhox5, Pramel7) in ES cells.
Main Results:
- Pluripotent, germline-competent ES cells were efficiently derived from STAT3-MER blastocysts without LIF, upon tamoxifen induction.
- Expression profiling identified 26 differentially expressed genes in tamoxifen-induced ES cells compared to wild-type.
- Four genes (Hexokinase II, Lefty2, Pramel7, PP1rs15B) showed inner cell mass (ICM)-restricted expression, indicating potential roles in pluripotency.
- Overexpression of Pem/Rhox5 and Pramel7 maintained ES cell markers and morphology independently of LIF.
Conclusions:
- STAT3 activation in the blastocyst ICM facilitates ES cell establishment and upregulates potential pluripotency maintenance genes.
- Pem/Rhox5 and Pramel7 are identified as key genes capable of maintaining ES cell pluripotency.
- These candidate genes can sustain ES cell pluripotency in a LIF-independent manner, similar to STAT3 and Nanog.
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