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Methods to Classify Cytoplasmic Foci as Mammalian Stress Granules
Published on: May 12, 2017
The Igf2as transcript is exported into cytoplasm and associated with polysomes.
Carolina Duart-Garcia1, Martin H Braunschweig
1Institute of Genetics, Vetsuisse Faculty, University of Bern, Bremgartenstrasse 109a, 3001, Berne, Switzerland.
Biochemical Genetics
|October 31, 2012
Summary
Murine insulin-like growth factor 2 antisense (Igf2as) transcripts are found in the cytoplasm and associate with polysomes. This suggests that Igf2as may function as a protein-coding transcript.
Area of Science:
- Genetics
- Molecular Biology
- Epigenetics
Background:
- Insulin-like growth factor 2 (Igf2) and its antisense transcript (Igf2as) are imprinted genes located on chromosome 7.
- Imprinting defects of IGF2 are linked to human developmental disorders and cancers.
- Understanding the function of Igf2as is crucial for comprehending Igf2 locus regulation.
Purpose of the Study:
- To investigate the cellular localization and potential function of murine Igf2as transcripts.
- To determine if Igf2as transcripts associate with the translational machinery.
Main Methods:
- Development of a RNA-FISH protocol to detect Igf2as and Igf2 transcripts.
- Quantitative real-time PCR for transcript validation.
- Polysome association studies using sucrose gradient centrifugation.
Main Results:
- RNA-FISH confirmed Igf2as transcripts are predominantly located in the cytoplasm of C2C12 cells.
- Quantitative real-time PCR validated RNA-FISH findings.
- Polysome fractionation analysis demonstrated that Igf2as transcripts sediment with polysomes.
Conclusions:
- The cytoplasmic localization and polysome association of Igf2as transcripts provide strong evidence for its role as a protein-coding molecule.
- Further studies are warranted to identify the protein product of Igf2as and its biological function.
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