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Abstract: TH-PO367

Identification of Circular RNAs Underlying Circadian Cycling in Mouse Kidney Samples

Session Information

Category: Cell Biology

  • 201 Cell Signaling, Oxidative Stress

Authors

  • Braun, Fabian, University Hospital Cologne, Cologne, NW, Germany
  • Johnsen, Marc, University Hospital Cologne, Cologne, NW, Germany
  • Schermer, Bernhard, University Hospital Cologne, Cologne, NW, Germany
  • Benzing, Thomas, University Hospital Cologne, Cologne, NW, Germany
  • Westermark, Pål O., Leibniz Institute for Farm Animal Biology, Dummerstorf, Germany
  • Mueller, Roman-Ulrich, University Hospital Cologne, Cologne, NW, Germany
Background

Over the past years circular RNAs have emerged as a new species of noncoding RNAs and a subject of intense research efforts. They are more stable than linear stranded RNA species and seem to be involved in post transcriptional gene regulation. Only last year, the first dataset of circular RNAs expressed in the murine kidney was published, yet we are far from understanding the precise involvement and bio molecular functions of these nucleic acids. Furthermore the question whether their expression is partially regulated in a timely or circadian manner has not been addressed so far.

Methods

10 week old wild type mice were sacrificed for their organs every three hours starting at 12pm for an overall period of 48 hours. We isolated RNA from the extracted kidneys, pooled samples of three mice per time point and performed RNA Sequencing. Using bioinformatics analyses of backsplicing events we identified circular RNA sequences. Furthermore, we analyzed quantitative changes of these backsplicing events during the circadian cycle. qPCR was used to further validate the rhythmicity of the cycling circular RNAs as well as their host genes.

Results

We identified more than 4000 distinct backsplicing events in our data set. A subset of these circles showed a distinct circadian rhythmicity. Of the 35 top cycling circular RNAs, we validated the circadian expression pattern for two circular RNAs derived from the Strn3 and Smad4 gene locus. Interestingly both host genes did not display a circadian expression pattern.

Conclusion

Our data set yields further insight into the expression pattern of circular RNAs in the murine kidney. Especially we present the first evidence for a circadian expression of certain circular RNA species. Ongoing experiments will focus on the analysis of potential miRNA binding sites on the two circadian circular RNAs and the rhythmicity of the potentially bound miRNAs.