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Abstract: FR-PO931

Insights from Genome-Wide MicroRNA Target Identification In Podocytes Using Argonaute PAR-CLIP

Session Information

Category: Glomerular Diseases

  • 1204 Podocyte Biology

Authors

  • Gur-Wahnon, Devorah, Hadassah Medical Center, Jerusalem, Israel
  • Navon, Royi, Hebrew University, Maale adumim, Israel
  • Ben-Dov, Iddo Z., Hadassah - Hebrew University Medical Center, Jerusalem, Israel

Group or Team Name

  • Laboratory of Medical Transcriptomics
Background

MicroRNAs (miRNA) are a class of small non-coding RNAs that regulate gene expression through inhibition of translation or stability of target mRNAs. While several studies have shown the crucial role played by miRNAs in podocyte development and homeostasis, the specific miRNA and miRNA targets remain under-explored. Quantitative miRNA profiling is essential, but has a limitation: the inhibitory activity of a miRNA depends not only on its absolute expression level in the cell, but also on its loading onto the RISC complex and the availability of its target binding sites.

Methods

We have therefore applied argonaute-2 (AGO2) PAR-CLIP (Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation) to provide us with the miRNA target principles in podocytes. We searched for enriched miRNA seed-complimentary regions within PAR-CLIP output clusters using an in-house algorithm as well as Sylamer.

Results

The most enriched 6-8-mer sequences complimentary to the seed of miRNA that are expressed in podocytes, show extensive overlap between human and mouse. Surprisingly, the top enriched sequences were complementary to the seed of miR-27a and miR-27b, which are not among the most abundant miRNAs in podocytes, and thus might have been overlooked based on miRNA profiling alone. Subsequently, knocking down miR-27 expression in podocytes resulted in their reduced survival as well as an altered actin cytoskeleton pattern.

Conclusion

These results suggest a central role for miR-27 in podocyte maintenance. Furthermore our work implies that researchers interested in miRNA research may apply similar methods when choosing miRNA and miRNA targets suitable for their research.

Funding

  • Government Support - Non-U.S.