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Related Experiment Videos

Protocol to study in vivo circRNA interactions in the mouse cortex using an RNA pull-down approach.

Valentina Silenzi1, Nicolò Salvi1, Irene Bozzoni2

  • 1Department of Biology and Biotechnologies "Charles Darwin," Sapienza University of Rome, 00185 Rome, Italy.

STAR Protocols
|September 19, 2025
PubMed
Summary
This summary is machine-generated.

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This study details a new protocol to identify circular RNA (circRNA) interactors in mouse brain tissue. The method uses UV crosslinking and RNA pull-down to find proteins associated with specific circRNAs.

Area of Science:

  • Molecular Biology
  • Neuroscience
  • Genomics

Background:

  • Circular RNAs (circRNAs) are a class of non-coding RNAs with emerging roles in gene regulation.
  • Identifying circRNA-binding proteins is crucial for understanding their functions.
  • Existing methods may have limitations in preserving native interactions.

Purpose of the Study:

  • To present a robust protocol for identifying protein interactors of circRNAs in the mouse cortex.
  • To optimize techniques for preserving native circRNA-protein complexes.
  • To provide a adaptable method for nervous system research.

Main Methods:

  • Tissue dissociation and UV crosslinking of mouse cortical tissue to stabilize RNA-protein interactions.
  • RNA pull-down assays using a specific circular RNA (circDlc1(2)) as bait.
Keywords:
Cell BiologyGene ExpressionMolecular BiologyNeuroscienceProtein Biochemistry

Related Experiment Videos

  • Optimization of steps for efficient isolation of circRNA-associated molecules.

    • Main Results:

    • The protocol successfully identifies potential protein interactors associated with circDlc1(2).
    • UV crosslinking effectively maintains native interactions between circRNAs and proteins.
    • The method is shown to be effective in mouse brain tissue.

    Conclusions:

    • This protocol provides a reliable method for discovering circRNA-protein interactions in the central nervous system.
    • The technique can be adapted for studying other circRNAs and brain regions.
    • Further research can utilize this protocol to elucidate circRNA functions in neurological processes.