Trans-spliced mRNA Products Produced from circRNA Expression Vectors
- 1 Dept of Biology, Massachussetts Institute of Technology, Cambridge, MA 02138, USA;
- 2 Department of Biochemistry, McGill University, Montreal, Canada;
- 3 Department of Biochemistry, Department of Oncology, Rosalind & Morris Goodman Cancer Research Centre, McGill University, Montreal, Canada
- ↵* Corresponding author; email: 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.
Keywords
- Received November 2, 2020.
- Accepted March 21, 2021.
- Published by Cold Spring Harbor Laboratory Press for the RNA Society
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