Abstract
Circular RNAs (circRNAs) are covalently closed transcripts generated by back-splicing reaction. The lack of free ends endows these RNA molecules with high stability thus allowing them to accumulate in tissues and body fluids. They are widely expressed in most organisms, are modulated during development and display tissue-specific expression, resulting particularly enriched in the nervous system. Deregulation of circRNA expression has also been associated with several pathological conditions including neurological diseases and cancer.
Here we present a Northern blot procedure that allows the analysis of the expression of bona fide circRNAs through the use of a digoxigenin-labeled RNA probe and the immunodetection of the signals.
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Acknowledgments
The authors would like to thank Lorenzo Errichelli and Ivano Legnini for the images in Fig. 3, panels c and d (reproduced respectively from ref. 25 which is licensed under the Creative Commons Attribution 4.0 International License and from ref. 16 with permission from Elsevier).
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D’Ambra, E., Morlando, M. (2021). Study of Circular RNA Expression by Nonradioactive Northern Blot Procedure. In: Navarro, A. (eds) Long Non-Coding RNAs in Cancer. Methods in Molecular Biology, vol 2348. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1581-2_23
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DOI: https://doi.org/10.1007/978-1-0716-1581-2_23
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