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Andreas W Heumüller1,2, Alisha N Jones3,4, André Mourão3,4

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Circular RNAs (circRNAs) regulate cellular functions. Locus-conserved circRNA cZNF292/cZfp292 impacts endothelial cell shape and flow responses by interacting with the protein SDOS.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Circular RNAs (circRNAs) are emerging regulatory RNAs with largely unknown in vivo functions.
  • Genetic inactivation of circRNAs is challenging, hindering studies on their physiological roles and conservation.
  • This study aimed to identify conserved circRNAs for functional analysis through genetic deletion.

Purpose of the Study:

  • To identify locus-conserved circRNAs between mice and humans.
  • To investigate the in vivo function of conserved circRNAs, specifically cZfp292.
  • To elucidate the molecular mechanism underlying circRNA-mediated endothelial cell regulation.

Main Methods:

  • Combined RNA-sequencing data with circRNA databases to identify conserved circRNAs.
  • Utilized CRISPR/Cas9 for genetic depletion of circRNA cZfp292 in vivo and in vitro.
  • Employed RNA-affinity purification and mass spectrometry to identify interacting proteins.
  • Investigated the role of protein SDOS and its binding partner Syndecan-4.

Main Results:

  • Identified locus-conserved circRNAs between mice and humans.
  • Genetic depletion of cZfp292 altered endothelial morphology and aortic flow alignment in vivo.
  • cZNF292 depletion in vitro abolished laminar flow-induced changes in endothelial cell orientation and focal adhesion.
  • Discovered specific interaction between cZNF292 and protein SDOS.
  • SDOS or Syndecan-4 silencing, or disruption of their binding site, recapitulated cZfp292 knockout phenotypes.

Conclusions:

  • Revealed a novel role for cZNF292/cZfp292 in endothelial cell responses to blood flow.
  • Demonstrated that cZNF292 influences endothelial cell shape and cytoskeletal organization.
  • Established a molecular mechanism involving SDOS and Syndecan-4 in circRNA-mediated flow responses.