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mRNA trans-splicing dual AAV vectors for (epi)genome editing and gene therapy

Author

Listed:
  • Lisa Maria Riedmayr

    (LMU Munich)

  • Klara Sonnie Hinrichsmeyer

    (LMU Munich)

  • Stefan Bernhard Thalhammer

    (LMU Munich)

  • David Manuel Mittas

    (LMU Munich)

  • Nina Karguth

    (LMU Munich)

  • Dina Yehia Otify

    (LMU Munich)

  • Sybille Böhm

    (ViGeneron GmbH)

  • Valentin Johannes Weber

    (University of Zurich)

  • Michael David Bartoschek

    (ViGeneron GmbH)

  • Victoria Splith

    (ViGeneron GmbH)

  • Manuela Brümmer

    (LMU Munich)

  • Raphael Ferreira

    (Harvard Medical School)

  • Nanda Boon

    (Leiden University Medical Center (LUMC))

  • Gabriele Maria Wögenstein

    (University of Zurich)

  • Christian Grimm

    (University of Zurich)

  • Jan Wijnholds

    (Leiden University Medical Center (LUMC)
    Institute of the Royal Netherlands Academy of Arts and Sciences (KNAW))

  • Verena Mehlfeld

    (LMU Munich)

  • Stylianos Michalakis

    (LMU Munich)

  • Stefanie Fenske

    (LMU Munich
    partner site Munich Heart Alliance)

  • Martin Biel

    (LMU Munich)

  • Elvir Becirovic

    (University of Zurich)

Abstract

Large genes including several CRISPR-Cas modules like gene activators (CRISPRa) require dual adeno-associated viral (AAV) vectors for an efficient in vivo delivery and expression. Current dual AAV vector approaches have important limitations, e.g., low reconstitution efficiency, production of alien proteins, or low flexibility in split site selection. Here, we present a dual AAV vector technology based on reconstitution via mRNA trans-splicing (REVeRT). REVeRT is flexible in split site selection and can efficiently reconstitute different split genes in numerous in vitro models, in human organoids, and in vivo. Furthermore, REVeRT can functionally reconstitute a CRISPRa module targeting genes in various mouse tissues and organs in single or multiplexed approaches upon different routes of administration. Finally, REVeRT enabled the reconstitution of full-length ABCA4 after intravitreal injection in a mouse model of Stargardt disease. Due to its flexibility and efficiency REVeRT harbors great potential for basic research and clinical applications.

Suggested Citation

  • Lisa Maria Riedmayr & Klara Sonnie Hinrichsmeyer & Stefan Bernhard Thalhammer & David Manuel Mittas & Nina Karguth & Dina Yehia Otify & Sybille Böhm & Valentin Johannes Weber & Michael David Bartosche, 2023. "mRNA trans-splicing dual AAV vectors for (epi)genome editing and gene therapy," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-42386-0
    DOI: 10.1038/s41467-023-42386-0
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    References listed on IDEAS

    as
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