CHARIOT-AAV: Conjugation of diverse vectors to adeno-associated viruses for delivery of large genes
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.
Authors
- Daniel G. Anderson (ORCID: https://orcid.org/0000-0001-5629-4798)
- Robert J. Macfarlane
- Tatsuya Osaki (ORCID: https://orcid.org/0000-0001-7174-0629)
- Chae Gyu Lee (ORCID: https://orcid.org/0000-0002-7887-5312)
- Jacob L. Beckham (ORCID: https://orcid.org/0000-0002-2787-3712)
- E. Guerra
- Keisuke Nagao (ORCID: https://orcid.org/0009-0002-2296-1817)
- Polina Anikeeva (ORCID: https://orcid.org/0000-0001-6495-5197)
- Emmanuel Vargas Paniagua
- Scott Machen
- Mriganka Sur
- Sho Toyonaga
Institutions
- McGovern Institute for Brain Research (US)
- Fujifilm (Japan) (JP)
- Institute of Cognitive and Brain Sciences (US)
- Whitehead Institute for Biomedical Research (US)
- Massachusetts Institute of Technology (US)
Publication Details
- Journal
- bioRxiv (Cold Spring Harbor Laboratory)
- Published
- 2026-04-15
- DOI
- https://doi.org/10.64898/2026.04.13.718054
- Primary Topic
- Virus-based gene therapy research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Science Foundation
- Simons Foundation
- Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology
- Fujifilm Corporation
- Honjo International Scholarship Foundation
- National Institutes of Health
- Materials Research Science and Engineering Center, Harvard University
- Multidisciplinary University Research Initiative
- Simons Center for the Social Brain, Massachusetts Institute of Technology
- McGovern Institute for Brain Research, Massachusetts Institute of Technology
- Division of Materials Research
- Army Research Office