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Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal
Published on: May 30, 2012
Myc-regulated microRNAs attenuate embryonic stem cell differentiation.
Chin-Hsing Lin1, Aimee L Jackson, Jie Guo
1Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-4417, USA.
The EMBO Journal
|September 12, 2009
Summary
The study reveals that the Myc protein regulates embryonic stem cell maintenance by controlling specific microRNAs (miRNAs). These miRNAs help prevent stem cell differentiation, maintaining pluripotency.
Area of Science:
- Stem cell biology
- Molecular and cellular biology
- Epigenetics and gene regulation
Background:
- Myc proteins are crucial for stem cell maintenance and regulate proliferation via microRNAs (miRNAs).
- The role of c-Myc in embryonic stem (ES) cell maintenance and differentiation through miRNAs remains to be fully elucidated.
Purpose of the Study:
- To investigate whether c-Myc influences ES cell maintenance and differentiation through the regulation of miRNAs.
- To identify specific miRNAs regulated by c-Myc in ES cells and their downstream targets.
Main Methods:
- Quantitative primer-extension PCR assay to identify miRNA expression.
- Chromatin immunoprecipitation to assess c-Myc binding near miRNA genes.
- Expression profiling and seed homology to identify miRNA and c-Myc targets.
- Introduction of c-Myc-induced miRNAs and knockdown of endogenous miRNAs in murine ES cells.
Main Results:
- c-Myc regulates specific miRNAs (e.g., miR-141, miR-200, miR-429) in ES cells, but not in differentiated derivatives.
- c-Myc binds to genomic regions encoding these miRNAs in ES cells.
- These miRNAs and c-Myc downregulate common target genes.
- Overexpression of c-Myc-induced miRNAs attenuates differentiation marker downregulation, while knockdown accelerates differentiation.
Conclusions:
- c-Myc plays a role in ES cell maintenance, partly through a subset of miRNAs.
- These miRNAs act to attenuate differentiation, thereby contributing to pluripotency maintenance.
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