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Initially, three interviewer researchers independently coded two interview transcripts (phase 1).
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For reference, Rev-erb β is a typical E-box gene that peaks at C10 and Bmal1 is a typical RRE regulated gene that peaks at CT0. Genes in this list regulated by both elements showed a spread of transcripts phases throughout the day with a bias towards dusk-phased E-box genes.
After miRNA guided cleavage of the soybean TAS3 transcripts, phased 21-nucleotide tasiRNAs are processed that, in turn, could target genes with auxin response factor motifs.
Since transcript phase is a circular dependent variable (i.e., phase CT24 is equal to CT0), we needed a link function to relate the linear predictor based on the three scores to the transcript phase.
One remarkable feature was that the transcript phase for at least the master list genes were quite consistent across 14 tissues.
We examined this hypothesis of whether binding of certain TFs was indicative of phase-specific transcript regulation using the ChIP-seq data on different circadian TFs and the transcript phase of master list genes.
However, transcript phases (time of peak expression) of robust candidate genes were strongly concentrated around CT9-13 (Fig. 3 a).
It must be borne in mind that for this analysis of phase regulation, we pooled all the transcript phases and assumed that ChIP-seq binding in the liver implies similar binding in all tissues.
It is important to note that p21 transcript phases are delayed in Per2 allele mutant MEFs, which may bring changes in the relative phases between p21 and Cyclin D. For the p21 gene, it has been described previously that its regulatory region contains a RORE binding site and is regulated by the antagonistic activities of the orphan nuclear receptors NR1D1/2 and NR1F1.
JB read all transcripts from phase two to verify the analysis.
As part of that process TSH reviewed and reanalyzed all the transcripts from phase two to strengthen the analysis.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com