Gene Therapy of mdx Mice With Large Truncated Dystrophins Generated by Recombination Using rAAV6
Open accessAbstract
Recombinant adeno-associated viral (rAAV) vector-mediated gene transfer represents a promising approach for many diseases. However, the applicability of rAAV vectors has long been hindered by the small (~4.8 kb) DNA packaging capacity. This limitation can hamper the packaging and delivery of critical regulatory elements and/or larger coding sequences, such as the ~14-kb dystrophin complementary DNA (cDNA) that is of interest for gene therapy of Duchenne muscular dystrophy (DMD). Here, we have demonstrated reconstitution of an expression cassette (7.3 kb) encoding a highly functional "minidystrophin" protein (ΔH2-R19, 222 kd) in vivo following intravascular co-delivery of two independent rAAV6 vectors sharing a central homologous recombinogenic region of 372 nucleotides. Similar to previously reported trans-splicing approaches, one rAAV vector provides the promoter with the ~1/2 initial portion of minidystrophin, while the second vector provides the remaining minidystrophin cDNA followed by the polyadenylation signal. Significantly, administering a modest dose [2 × 10(12) vector genomes (vg)] of the two minidystrophin-encoding rAAV vectors to dystrophic mice elicited an improvement of physiological performance indicative of prevention or amelioration of the disease state. These studies provide evidence that functional dystrophin transgenes larger than that typically carried by a single rAAV genome can be reconstituted in vivo by homologous recombination (HR) following intravas…
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Overlapping vectors work particularly well in muscle tissue, ameliorating phenotypes linked to Duchenne muscular dystrophy and dysferlinopathies.
“This strategy was shown to work particularly well in muscle tissue, ameliorating phenotypes linked to Duchenne muscular dystrophy (DMD) [43][44][45] and dysferlinopathies [46][47][48].”
From Circumventing the packaging limit of AAV-mediated gene replacement therapy for neurological disorders (2022) · cites this paper as [43]
The microdystrophin variant can be packaged in a single AAV vector.
“the so-called microdystrophin variant can be packaged in a single AAV vector”
From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [42]
In mdx mice, gene transfer with AAV vectors encoding microdystrophin provides significant, although not optimal, therapeutic benefit.
“in mdx mice (a mouse model of Duchenne muscular dystrophy) gene transfer with AAV vectors encoding microdystrophin provides significant, although not optimal therapeutic benefit. 40-42”
From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [42]
The overlap method was used successfully with a split mini-dystrophin expression cassette with a combined length of approximately 7.3 kb and a short 372 bp overlap region.
“the overlap method was also used successfully with a split mini-dystrophin expression cassette with a combined length of approximately 7.3 kb and a short, 372 bp overlap region. 42”
From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [42]
Expression of mini-dystrophin was observed in striated muscle of mdx mice when rAAV6 was given intravenously.
“expression of mini-dystrophin was observed in striated muscled of mdx-mice when rAAV6 was given intravenously.”
From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [42]
Although overall levels of mini-dystrophin expression were lower than in wildtype muscles, there was an improvement in limb muscle physiological performance.
“Although the overall levels of minidystrophin expression were lower than in wildtype muscles, there was an improvement in limb muscle physiological performance. 42”
From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [42]
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Bengtsson, N. E., Hall, J. K., Odom, G. L., Phelps, M. P., Andrus, C. R., Hawkins, R. D., Hauschka, S. D., Chamberlain, J. R., & Chamberlain, J. S. (2017) · Nature Communications · doi:10.1038/ncomms14454 · Cited by 387 · see citing works
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- Authors
- Odom, G. L., Gregorevic, P., Allen, J. M., & Chamberlain, J. S.
- Affiliations
- University of Washington (US)
- Corresponding author
- Chamberlain, Jeffrey S
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2011-01
- Volume
- 19
- Issue
- 1
- Pages
- 36-45
- ISSN
- 1525-0016 (ISSN-L)
- PMC
- PMC3017440
- OpenAlex
- W2147735923
- Cited by
- 95
- References
- 6
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 5 · 2023: 4 · 2022: 3 · 2021: 1 · 2020: 3 · 2019: 6 · 2018: 2 · 2017: 11 · 2016: 4 · 2015: 15 · 2014: 10 · 2013: 9 · 2012: 12
- FWCI
- 4.86
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Muscular Dystrophy Association
- National Institute for Health and Care Research
- National Institutes of Health
- National Institute on Aging (AG033610, R01AG033610)
- National Heart, Lung, and Blood Institute (T32-HL-07828, T32HL007828)
- National Institute of Arthritis and Musculoskeletal and Skin Diseases (R37AR040864, AR 40864, R01AR040864)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Health Sciences › Medicine › Genetics › Neurogenetic and Muscular Disorders Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- Duchenne muscular dystrophy, dystrophin, recombinant adeno-associated virus vectors, homologous recombination, trans-splicing, mdx mice, gene therapy
- MeSH terms
- Dependovirus, Animals, Genetic Vectors, Injections, Intravenous, Male, Peptide Fragments, Recombination, Genetic, Genetic Therapy, Dystrophin, Open Reading Frames, Genome, Viral, Gene Transfer Techniques, DNA, Complementary, Mice, Inbred mdx, Muscle, Skeletal, Transgenes, Trans-Splicing, Muscular Dystrophy, Duchenne, DNA Packaging, Mice
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