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Best practices to ensure robust investigation of circular RNAs: pitfalls and tips
Author(s) -
Dodbele Samantha,
Mutlu Nebibe,
Wilusz Jeremy E
Publication year - 2021
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.15252/embr.202052072
Subject(s) - microrna , computational biology , biology , circular rna , rna , translation (biology) , identification (biology) , function (biology) , clarity , non coding rna , gene , messenger rna , genetics , microbiology and biotechnology , botany
Pre‐mRNAs from thousands of eukaryotic genes can be non‐canonically spliced to generate circular RNAs (circRNAs) that have covalently linked ends. Most mature circular RNAs are expressed at low levels, but some have known physiological functions and/or accumulate to higher levels than their associated linear mRNAs. These observations have sparked great interest into this class of previously underappreciated RNAs and prompted the development of new experimental approaches to study them, especially methods to measure or modulate circular RNA expression levels. Nonetheless, each of these approaches has caveats and potential pitfalls that must be controlled for when designing experiments and interpreting results. Here, we provide practical advice, tips, and suggested guidelines for performing robust identification, validation, and functional characterization of circular RNAs. Beyond promoting rigor and reproducibility, these suggestions should help bring clarity to the field, especially how circular RNAs function and whether these transcripts may sponge microRNAs/proteins or serve as templates for translation.