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Using transcript-specific PCR assays, we found expression of both long and short transcripts of DCLK1 in the fetal brain (26 40 weeks) as well as in specific regions of the adult human brain (see Figure 1C).
Considering that one of the main signals of association was observed for markers in the intron 5 of DCLK1 and that short transcripts of DCLK1 all start from the exon 6, we hypothesised that the intron 5 might harbour alternative promoter and regulatory regions.
Prokaryotic asRNAs are generally short transcripts of approximately 100 to 300 nt [ 2].
Tags aligning to distinct long and short transcripts of FCA (cDNA clones are ~6.5 kb apart – Figure 2B) support this interpretation.
A technical feature of using high-throughput sequencing to interrogate full length transcripts is that longer transcripts produce more reads relative to short transcripts of similar expression.
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Another TSG candidate highlighted is the short transcript of SMAD6, variant 2, but not the full-length SMAD6, variant 1. SMAD transcription factors lie at the core of the transforming growth factor-beta (TGF-β) signaling pathway.
Intriguingly it was recently shown that in HD a short transcript of the HTT gene is produced by aberrant splicing, probably influenced by abnormal binding of the splicing factor SRSF6 to the CAG repeat expansion [ 19].
It should be noted that we detected shorter transcripts (of approximate 2 kb) for each gene in testes although the nature of these transcripts was not determined.
The reviewers feel the discovery that RNA Pol IV creates short transcripts is of major interest, as it substantially alters the existing model of RNA-directed DNA methylation.
Using real-time PCR, we report that the shorter transcript of SRRM2 was 1.7 fold (p = 0.008) upregulated in the substantia nigra of PDs vs controls while the longer transcript was 0.4 downregulated in both the substantia nigra (p = 0.03) and amygdala (p = 0.003).
Using real-time PCR on postmortem substantia nigra (SN) and amygdala from 10 PD patients versus 10 controls, we found that the shorter transcript of SRRM2 was significantly upregulated (FC = 1.7, p = 0.008) in the SN (as in the microarrays) but unchanged in the amygdala (AMG) of PD patients.
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