Trans-spliced mRNA products produced from circRNA expression vectors
- 1Department of Biochemistry, McGill University, Montreal, Canada, H3G 1Y6
- 2Department of Oncology, McGill University, Montreal, Canada, H3A 1G5
- 3Rosalind & Morris Goodman Cancer Research Centre, McGill University, Montreal, Canada, H3A 1A3
- 4Centre de Recherche en Biologie Structurale, McGill University, Montreal, Canada, H3G 1Y6
- Corresponding author: jerry.pelletier{at}mcgill.ca
Abstract
Circular (circ) RNA expression vectors are used as a method of identifying and characterizing RNA sequences that harbor internal ribosome entry site (IRES) activity. During the course of developing a vector series tailored for IRES discovery, we found evidence for the occurrence of trans-spliced mRNAs arising when sequences with promoter activity were embedded between the upstream CTD and downstream NTD exons of the pre-mRNA. These trans-spliced products regenerate the same open reading frame expected from a circRNA and can lead to false-positive signals in screens relying on circRNA expression vectors for IRES discovery. Our results caution against interpretations of IRES activity solely based on results obtained from circRNA expression vectors.
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Article is online at http://www.rnajournal.org/cgi/doi/10.1261/rna.078261.120.
- Received November 2, 2020.
- Accepted March 21, 2021.
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