Tissue-Dependent Expression and Translation of Circular RNAs with Recombinant AAV Vectors In Vivo

9Citations
Citations of this article
47Readers
Mendeley users who have this article in their library.

Abstract

Circular RNAs (circRNAs) are long-lived, covalently closed RNAs that are abundantly expressed and evolutionarily conserved across eukaryotes. Possible functions ranging from microRNA (miRNA) and RNA binding protein sponges to regulators of transcription and translation have been proposed. Here we describe the design and characterization of recombinant adeno-associated viral (AAV) vectors packaging transgene cassettes containing intronic sequences that promote backsplicing to generate circularized RNA transcripts. Using a split GFP transgene, we demonstrate the capacity of vectors containing different flanking intronic sequences to efficiently drive persistent circRNA formation in vitro. Further, translation from circRNA is efficiently driven by an internal ribosomal entry site (IRES). Upon injecting AAV vectors encoding circRNA in mice, we observed robust transgene expression in the heart, but low transduction in the liver for the intronic elements tested. Expression in the murine brain was restricted to astrocytes following systemic or intracranial administration, while intravitreal injection in the eye yielded robust transgene expression across multiple retinal cell layers. These results highlight the potential for exploiting AAV-based circRNA expression to study circRNA function and tissue-specific regulation in animal models, as well as development of therapeutic platforms using this approach.

Cite

CITATION STYLE

APA

Meganck, R. M., Borchardt, E. K., Castellanos Rivera, R. M., Scalabrino, M. L., Wilusz, J. E., Marzluff, W. F., & Asokan, A. (2018). Tissue-Dependent Expression and Translation of Circular RNAs with Recombinant AAV Vectors In Vivo. Molecular Therapy - Nucleic Acids, 13, 89–98. https://doi.org/10.1016/j.omtn.2018.08.008

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free