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Encapsulating In Vitro Transcribed circRNA into Lipid Nanoparticles Via Microfluidic Mixing

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Circular RNAs

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

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Abstract

This chapter serves as a guide for researchers embarking on circular RNA-based translational studies. It provides a foundation for the successful encapsulation of circular RNA into lipid nanoparticles (LNPs) and facilitates progress in this emerging field. Crucial scientific methods and techniques involved in the formulation process, particle characterization, and downstream processing of circ-LNPs are covered. The production of in vitro transcribed circular RNA-containing LNPs based on a commercially available lipid mix is provided, in addition to the fundamentals for successful encapsulation based on lipid mixes composed of single components. Furthermore, the transfection and validation protocols for the identification of a functional and potentially therapeutic circRNA candidate for initial in vitro verification, before subsequent LNP studies, are explained.

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Acknowledgments

This work was supported by funding from the German Research Foundation, DFG (SFB/Transregio TRR267, to C.B. and T.T.) and the European Research Council, ERC (ERC Advanced Grant REVERSE, to T.T.).

Disclosure

TT is a founder and shareholder of Cardior Pharmaceuticals GmbH (outside of this book chapter). D.L., C.B., and T.T. have filed and partly licensed patents for non-coding RNAs including for Circ-INSR.

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Author notes

  1. Malte Juchem and Sarah Cushman contributed equally.

Authors and Affiliations

  1. Institute of Molecular and Translational Therapeutic Strategies (IMTTS), Hannover Medical School, Hannover, Germany

    Malte Juchem, Sarah Cushman, Dongchao Lu, Shambhabi Chatterjee, Christian Bär & Thomas Thum

  2. Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Hannover, Germany

    Malte Juchem, Shambhabi Chatterjee & Christian Bär

Authors
  1. Malte Juchem
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  2. Sarah Cushman
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  3. Dongchao Lu
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  4. Shambhabi Chatterjee
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  5. Christian Bär
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  6. Thomas Thum
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Corresponding authors

Correspondence to Christian Bär or Thomas Thum .

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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Juchem, M., Cushman, S., Lu, D., Chatterjee, S., Bär, C., Thum, T. (2024). Encapsulating In Vitro Transcribed circRNA into Lipid Nanoparticles Via Microfluidic Mixing. 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_14

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  • DOI: https://doi.org/10.1007/978-1-0716-3678-7_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3677-0

  • Online ISBN: 978-1-0716-3678-7

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