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bioRxiv · 10.64898/2026.04.13.718054

CHARIOT-AAV: Conjugation of diverse vectors to adeno-associated viruses for delivery of large genes

Nagao, K.·Osaki, T.·Toyonaga, S.·Lee, C. G.·Vargas Paniagua, E.·Crespin Guerra, E.·Machen, S.·Beckham, J. L.·Sur, M.·Anderson, D. G.·Macfarlane, R. J.·Anikeeva, P.
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Abstract

Systemic, tissue-specific delivery of large transgenes exceeding the packaging capacity of adeno-associated viruses (AAVs) remains a key translational challenge for molecular therapeutics. Vectors with larger capacities, such as lentiviral vectors (LVVs) and lipid nanoparticles (LNPs), often lack adjustable, tissue-specific tropisms. Here we report CHARIOT-AAV (Crosslinked Hybrid Architectures for Robust, Interchangeable, and Organ-specific Targeting with AAV), a platform where diverse delivery vectors are conjugated to AAVs, thereby achieving tissue-specific tropism of AAVs and expanded cargo capacity. AAV-AAV conjugates packaging split SpCas9 constructs in AAV.CAP-B10 capsids demonstrate a [~]2-fold increase in brain gene editing efficiency over unconjugated AAV cocktails after intravenous injection. In addition to AAV-AAV conjugates, AAV-LVV and AAV-LNP conjugates achieve AAV-guided delivery of genetic payloads to target cells. Furthermore, AAV-LNP conjugates enable systemic delivery of mRNAs to brain endothelial cells. CHARIOT-AAV thus provides a modular platform for systemic, tissue-specific delivery of diverse therapeutics beyond the limits of individual vectors.

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Nagao, K., Osaki, T., Toyonaga, S., Lee, C. G., Vargas Paniagua, E., Crespin Guerra, E., Machen, S., Beckham, J. L., Sur, M., Anderson, D. G., Macfarlane, R. J., Anikeeva, P.. 2026-04-15. CHARIOT-AAV: Conjugation of diverse vectors to adeno-associated viruses for delivery of large genes. https://doi.org/10.64898/2026.04.13.718054

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