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Engineering Synthetic circRNAs for Efficient CNS Expression

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Circular RNAs

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2765))

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Abstract

Circular RNAs (circRNAs) have recently emerged as a promising modality for gene and RNA-based therapies. They are more stable than their linear counterpart and can be designed for efficient expression in different cell and tissue types. In this chapter, we developed different backsplicing circRNA cassettes that can enable efficient gene expression in various cell and tissue types. Furthermore, we packaged cassettes encoding circRNAs into adeno-associated viral (AAV) vectors that can be delivered via intracerebroventricular (ICV) injections to achieve expression in murine brain tissue. We provide detailed methods for the design of backsplicing circRNAs, circRNA detection, and generation of AAV-circRNA vectors for CNS dosing and expression in mice.

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Acknowledgments

This work was supported by the National Institutes of Health (R01NS099371 to A.A., W.F.M., and J.E.W. and R01HL089221, R01GM127708, and UG3AR075336 to A.A).

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Authors and Affiliations

  1. Department of Surgery, Duke University School of Medicine, Durham, NC, USA

    Katie N. Clements & Aravind Asokan

  2. Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, NC, USA

    Trevor J. Gonzalez & Aravind Asokan

  3. Department of Biomedical Engineering, Duke University, Durham, NC, USA

    Aravind Asokan

Authors
  1. Katie N. Clements
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  2. Trevor J. Gonzalez
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  3. Aravind Asokan
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Corresponding author

Correspondence to Aravind Asokan .

Editor information

Editors and Affiliations

  1. Klaus Tschira Institute for Integrative Computational Cardiology, University Heidelberg, Heidelberg, Germany

    Christoph Dieterich

  2. Department of Cellular, Computational and Integrative Biology – CIBIO, University of Trento, Trento, Italy

    Marie-Laure Baudet

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Clements, K.N., Gonzalez, T.J., Asokan, A. (2024). Engineering Synthetic circRNAs for Efficient CNS Expression. In: Dieterich, C., Baudet, ML. (eds) Circular RNAs. Methods in Molecular Biology, vol 2765. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3678-7_13

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  • DOI: https://doi.org/10.1007/978-1-0716-3678-7_13

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3677-0

  • Online ISBN: 978-1-0716-3678-7

  • eBook Packages: Springer Protocols

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