Journal
BIOCHEMICAL GENETICS
Volume 51, Issue 1-2, Pages 119-130Publisher
SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s10528-012-9547-8
Keywords
Igf2as; RNA-FISH; C2C12 cells; Genomic imprinting; Polysome analysis
Funding
- Swiss National Foundation [31003A_127564]
- Swiss National Science Foundation (SNF) [31003A_127564] Funding Source: Swiss National Science Foundation (SNF)
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Murine insulin-like growth factor 2 antisense (Igf2as) transcripts originate from the opposite strand of the same Igf2 locus as the Igf2 sense mRNA. The Igf2, insulin 2 (Ins2), and H19 genes form a cluster of imprinted genes on chromosome 7. Loss of imprinting of IGF2 in humans is associated with Beckwith-Wiedemann syndrome and Silver-Russell syndrome, as well as with Wilm's tumor and colorectal cancer. We developed a RNA-FISH protocol to detect Igf2as and Igf2 transcripts. The results from the RNA-FISH were confirmed with quantitative real-time PCR and clearly indicate that the Igf2as transcripts are predominantly located in the cytoplasm of C2C12 cells. In a polysome association study, we showed that the Igf2as sedimented with polysomes in a sucrose gradient. The cellular localization of Igf2as transcripts together with polysome fractionation analysis provides compelling evidence that the Igf2as is protein coding.
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