Efficient in vivo gene expression by trans-splicing adeno-associated viral vectors
Open accessAbstract
Although adeno-associated virus (AAV)-mediated gene therapy has been hindered by the small viral packaging capacity of the vector, trans-splicing AAV vectors are able to package twice the size of the vector genome. Unfortunately, the efficiency of current trans-splicing vectors is very low. Here we show that rational design of the gene splitting site has a profound influence on trans-splicing vector-mediated gene expression. Using mRNA accumulation as a guide, we generated a set of efficient trans-splicing vectors and achieved widespread expression of the 6-kb DeltaH2-R19 mini-dystrophin gene in skeletal muscle of mdx mice, a model for Duchenne muscular dystrophy. The dystrophic phenotype was ameliorated in both adult and aged mice. This demonstrates the use of trans-splicing vectors to efficiently express a large therapeutic structural protein. This strategy should be applicable to other large therapeutic genes or large transcription regulatory elements.
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Efficient trans-splicing relies on the chosen sequences, making it crucial to optimize vector design on a case-to-case basis.
“Importantly, it was also observed that particular care should be put into selecting a splitting point and splicing signals, as efficient trans-splicing relies on the chosen sequences, making it crucial to meticulously optimize vector design on a case-to-case basis [41].”
From Circumventing the packaging limit of AAV-mediated gene replacement therapy for neurological disorders (2022) · cites this paper as [41]
With the optimal exon/intron/exon combination, approximately 80% of myofibers expressed mini-dystrophin.
“with the optimal exon/intron/exon combination approximately 80% of myofibers expressed mini-dystrophin. 61”
From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [61]
The nature of the splice site substantially influences transduction efficiencies.
“Intramuscular injection of AAV6 vectors with different exon/intron/ exon splitting sites demonstrated that the nature of the splice site substantially influences transduction efficiencies”
From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [61]
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“to meticulously optimize vector design on a case-to-case basis [41].”
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- Authors
- Lai, Y., Yue, Y., Liu, M., Ghosh, A., Engelhardt, J. F., Chamberlain, J. S., & Duan, D.
- Affiliations
- University of Missouri (US)
- University of Iowa (US)
- University of Washington (US)
- Corresponding author
- Duan, Dongsheng
- Venue
- Nature Biotechnology
- Published
- 2005-10-09
- Volume
- 23
- Issue
- 11
- Pages
- 1435-1439
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- PMC
- PMC2581721
- OpenAlex
- W2053849856
- Cited by
- 224
- References
- 2
- Citations by year
- 2026: 5 · 2025: 9 · 2024: 10 · 2023: 7 · 2022: 9 · 2021: 9 · 2020: 11 · 2019: 12 · 2018: 8 · 2017: 10 · 2016: 9 · 2015: 13 · 2014: 10 · 2013: 8 · 2012: 7
- FWCI
- 8.27
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of Arthritis and Musculoskeletal and Skin Diseases (AR49419, R01AR049419)
- 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
- trans-splicing, adeno-associated virus vectors, in vivo gene expression, mini-dystrophin, Duchenne muscular dystrophy, mdx mice, skeletal muscle, gene therapy
- MeSH terms
- Dependovirus, Animals, Cell Line, Exons, Genetic Vectors, Humans, Introns, Male, Muscles, Phenotype, RNA Splicing, RNA, Messenger, Transcription, Genetic, Models, Statistical, Genetic Therapy, Dystrophin, Gene Transfer Techniques, Mice, Inbred mdx, Muscle, Skeletal, Trans-Splicing, Mice
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