Hybrid Adeno-Associated Virus Bearing Nonhomologous Inverted Terminal Repeats Enhances Dual-Vector Reconstruction of Minigenes In Vivo
Open accessAbstract
We have previously demonstrated that hybrid adeno-associated viral (AAV) vectors bearing nonhomologous inverted terminal repeats (ITRs) enhance directional intermolecular recombination and the efficiency of dual-AAV vector trans-splicing in cultured cells. Using hybrid-ITR vectors carrying two exons of a lacZ minigene, we demonstrate that this dual-vector approach also mediates higher levels (3- to 6-fold) of gene reconstitution in mouse skeletal muscle, liver, and heart. Inhibition of the proteasome by systemic administration of Doxil (Food and Drug Administration-approved lipid-formulated doxorubicin) further enhanced dual-vector trans-splicing 6- to 12-fold in two mouse strains. Hence, using hybrid-ITR AAV vectors in combination with proteasome modulation enhanced dual-vector delivery of a transgene approximately 36-fold over the current dual-vector trans-splicing approaches. These data provide in vivo evidence that ITR sequence-dependent homologous recombination, rather than nonhomologous end joining, is the predominant mechanism for AAV genome heterodimerization. Hence, enhanced directional recombination provided by hybrid-ITR vectors may be a useful in vivo strategy for improving dual-vector delivery of transgenes larger than the AAV packaging limit.
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AAV genomes can concatamerize in incorrect orientation, which can be at least partly counteracted using heterologous ITRs.
“Unfortunately, subsequent studies highlighted the possibility for AAV genomes to concatamerize in the incorrect orientation [39], which could be at least partly counteracted using heterologous ITRs (derived from different AAV serotypes) [40].”
From Circumventing the packaging limit of AAV-mediated gene replacement therapy for neurological disorders (2022) · cites this paper as [40]
Intramuscular injection of a vector pair with heterologous ITRs resulted in a 3-6-fold higher expression level compared with a homologous vector pair.
“intramuscular injection of a vector pair with heterologous ITRs resulted in a 3-6-fold higher expression level when compared with a homologous vector pair. 49,59”
From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [59]
Cites
Trans-splicing vectors expand the utility of adeno-associated virus for gene therapy
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Self-citationYan, Z., Zak, R., Zhang, Y., Ding, W., Godwin, S., Munson, K., Peluso, R., & Engelhardt, J. F. (2004) · Journal of Virology · doi:10.1128/jvi.78.6.2863-2874.2004 · Cited by 138 · see citing works
Self-citationNakai, H., Montini, E., Fuess, S., Storm, T. A., Meuse, L., Finegold, M., Grompe, M., & Kay, M. A. (2003) · Molecular Therapy · doi:10.1016/s1525-0016(02)00023-0 · Cited by 52 · see citing works
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Effective delivery of large genes to the retina by dual AAV vectors
Trapani, I., Colella, P., Sommella, A., Iodice, C., Cesi, G., de Simone, S., Marrocco, E., Rossi, S., Giunti, M., Palfi, A., et al. (2014) · EMBO Molecular Medicine · doi:10.1002/emmm.201302948 · Cited by 269 · see citing works
Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids
Chamberlain, K., Riyad, J. M., & Weber, T. (2016) · Human Gene Therapy Methods · doi:10.1089/hgtb.2015.140 · Cited by 141 · see citing works
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Adeno-Associated Viral Vectors as a Tool for Large Gene Delivery to the Retina
Trapani, I. (2019) · Genes · doi:10.3390/genes10040287 · Cited by 73 · see citing works
Marrone, L., Marchi, P. M., & Azzouz, M. (2022) · Expert Opinion on Biological Therapy · doi:10.1080/14712598.2022.2012148 · Cited by 56 · see citing works
“(derived from different AAV serotypes) [40].”
- Authors
- Yan, Z., Lei-Butters, D. C. M., Zhang, Y., Zak, R., & Engelhardt, J. F.
- Affiliations
- University of Iowa (US)
- Department of Anatomy and Cell Biology, University of Iowa School of Medicine, Iowa City, IA 52242.
- Venue
- Human Gene Therapy
- Publisher
- SAGE Publications
- Published
- 2007-01
- Volume
- 18
- Issue
- 1
- Pages
- 81-87
- ISSN
- 1043-0342 (ISSN-L)
- 1557-7422
- PMC
- PMC2121583
- OpenAlex
- W2008971344
- Cited by
- 41
- References
- 3
- Citations by year
- 2025: 2 · 2024: 1 · 2023: 2 · 2022: 1 · 2021: 3 · 2019: 5 · 2018: 1 · 2017: 3 · 2016: 3 · 2015: 3 · 2014: 2 · 2013: 3
- FWCI
- 1.78
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Human Gene Therapy (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Heart, Lung, and Blood Institute (R01HL058340, R01 HL58340)
- National Institute of Diabetes and Digestive and Kidney Diseases (DK54759, P30DK054759)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Health Sciences › Medicine › Genetics › Neurogenetic and Muscular Disorders Research
- Keywords
- adeno-associated virus vectors, gene reconstitution, homologous recombination, non-homologous end joining, proteasome inhibition, doxorubicin, pegylated liposomal doxorubicin, transgene delivery, skeletal muscle, liver, heart
- MeSH terms
- Dependovirus, Animals, Antibiotics, Antineoplastic, Cell Line, Doxorubicin, Genetic Vectors, Lac Operon, Transduction, Genetic, Gene Expression Regulation, Viral, Genome, Viral, Transgenes, Trans-Splicing, Terminal Repeat Sequences, Mice
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