Abstract
Circular RNAs (circRNAs) are a widespread, cell-, tissue-, and disease-specific class of largely non-coding RNA transcripts. These single-stranded, covalently-closed transcripts arise through non-canonical splicing of pre-mRNA, a process called back-splicing. Back-splicing results in circRNAs which are distinguishable from their cognate mRNA as they possess a unique sequence of nucleic acids called the backsplice junction (BSJ). CircRNAs have been shown to play key functional roles in various cellular contexts and achieve this through their interaction with other macromolecules, particularly other RNA molecules and proteins. To elucidate the molecular mechanisms underlying circRNA function, it is necessary to identify these interacting partners. Herein, we present an optimized strategy for the simultaneous purification of the circRNA interactome within eukaryotic cells, allowing the identification of both circRNA–RNA and circRNA–protein interactions.
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Acknowledgments
Research reported in this publication was supported by the National Health and Medical Research Council (NHMRC) project grant funding to S.J.C. (GNT1144250). Fellowship support was provided by the Australian Research Council Future Fellowship to S.J.C. (FT160100318) and the NHMRC Investigator Leadership Grant to S.J.C. (GNT1198014). Fellowship support for B.W.S. was provided by the Flinders Foundation Brain Cancer Fellowship. We would like to acknowledge Flinders Proteomics for their guidance and expertise in regards to mass spectrometry methodologies.
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Gabryelska, M.M. et al. (2024). Native Circular RNA Pulldown Method to Simultaneously Profile RNA and Protein Interactions. In: Dieterich, C., Baudet, ML. (eds) Circular RNAs. Methods in Molecular Biology, vol 2765. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3678-7_16
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DOI: https://doi.org/10.1007/978-1-0716-3678-7_16
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