Efficient trans-splicing in the retina expands the utility of adeno-associated virus as a vector for gene therapy
Abstract
Recombinant vectors based on adeno-associated virus (AAV) can efficiently transduce many different cell types, including cells of the retina, resulting in stable gene expression. A major shortcoming of this vector is its small packaging capacity. A trans-splicing approach, which reconstitutes gene expression from two independent AAV vectors, can be used to overcome the vector's packaging limitations. The efficiency of this system to date has been disappointing, and therefore its utility for therapeutic application limited. We demonstrate here that efficiency and cellular specificity of trans-splicing is dependent on selection of the appropriate AAV serotype. Efficiency of transgene expression resulting from trans-splicing in skeletal muscle approaches that obtained when delivering the intact transgene when using AAV2 vectors packaged with AAV5 capsids (AAV2/5). This expands the potential of AAV vectors for retinal gene therapy. The use of AAV2/5 also increases the efficiency of trans-splicing in photoreceptors. Selection of the appropriate AAV serotype is likely to affect efficiency of trans-splicing in other organ systems as well.
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In the retina, the dual approach was first reported by Reich et al. (2003) using trans-splicing to show β-galactosidase expression in vivo around 40% after AAV5 subretinal injection.
“In the retina, the dual approach was first reported by Reich et al. (2003) which used the trans-splice technique to show that β-galactosidase expression in vivo was around 40% after AAV5 subretinal injection (Reich et al., 2003).”
From Evaluating Efficiencies of Dual AAV Approaches for Retinal Targeting (2017) · cites this paper as Reich et al., 2003
The trans-splicing method to expand the size of ORFs that can be successfully expressed using AAV vectors was originally developed in the laboratory of John Engelhardt and has since been utilized in several studies with varying success.
“The trans-splicing method to expand the size of ORFs that can be successfully expressed using AAV vectors was originally developed in the laboratory of John Engelhardt 52,53 and has since been utilized in several studies with varying success (e.g., refs. 53-57 ).”
From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [56]
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Trans-splicing vectors expand the utility of adeno-associated virus for gene therapy
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- Authors
- Reich, S. J., Auricchio, A., Hildinger, M., Glover, E., Maguire, A. M., Wilson, J. M., & Bennett, J.
- Affiliations
- F.M. Kirby Center for Molecular Ophthalmology, Department of Ophthalmology, Scheie Eye Institute, Philadelphia, PA 19104
- Telethon Institute Of Genetics And Medicine (IT)
- The Wistar Institute (US)
- Venue
- Human Gene Therapy
- Publisher
- SAGE Publications
- Published
- 2003-01
- Volume
- 14
- Issue
- 1
- Pages
- 37-44
- ISSN
- 1043-0342 (ISSN-L)
- 1557-7422
- OpenAlex
- W2152509882
- Cited by
- 96
- References
- 4
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 2 · 2023: 2 · 2021: 2 · 2020: 5 · 2019: 4 · 2018: 2 · 2017: 5 · 2016: 2 · 2015: 10 · 2014: 4 · 2013: 3 · 2012: 5
- FWCI
- 4.24
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Human Gene Therapy (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Eye Institute (EY10820, EY-12156, R01EY010820, R01EY012156)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- trans-splicing, adeno-associated virus vectors, retinal gene therapy, AAV2/5, photoreceptors, skeletal muscle, transgene expression efficiency
- MeSH terms
- Dependovirus, Animals, Base Sequence, beta-Galactosidase, Capsid, Cells, Cultured, Gene Expression Regulation, Genetic Vectors, Humans, Mice, Inbred Strains, Molecular Sequence Data, Organ Specificity, Retina, Serotyping, Genetic Therapy, Muscle, Skeletal, Transgenes, Trans-Splicing, Mice
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