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Time Course of Transgene Expression After Intrastriatal Pseudotyped rAAV2/1, rAAV2/2, rAAV2/5, and rAAV2/8 Transduction in the Rat
Open accessAbstract
In vivo recombinant adeno-associated viral vector (rAAV)-mediated transduction of various tissues including brain has been characterized by slow onset and gradual increase in gene expression before reaching stable long-term protein levels. The early time course of transgene expression has not been quantified using newly available rAAV capsid serotypes. In this experiment, the onset of expression of green fluorescent protein (GFP) after intrastriatal injection of rAAV2-based pseudotyped vectors (rAAV1, rAAV5, and rAAV8 capsids) was quantified. Native GFP fluorescence displayed a delayed onset of expression of at least 7 days for all the pseudotyped rAAV vectors. However, GFP immunohistochemical staining revealed significant transgene expression by 4 days after transduction for all serotypes and stable GFP(+) neuronal populations mediated by all serotypes within 14 days post transduction at the latest. rAAV2/1 and rAAV2/2 displayed no time-dependent increase of GFP(+) striatal neurons; reaching maximal striatal cell GFP(+) counts at 4 days after injection. All serotypes displayed peak transgene expression by 4 weeks post injection where native GFP(+) neurons were equal to immunostained striatal GFP(+) neurons. The inflammatory response to these rAAV vectors was present up to 4 weeks after transduction but was not apparent 9 months post injection. Thus, rAAV-mediated transgene expression begins earlier than previously thought.
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“In our previous work, we have seen that a capsid and/or transgene related inflammatory response peaks at 2-weeks post-injection and then retracts to the needle track [46], similarly to what is seen with the Olig001-GFP injected monkeys.”
From Novel oligodendroglial alpha synuclein viral vector models of multiple system atrophy: studies in rodents and nonhuman primates (2017) · cites this paper as [46]
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Metadata
- Authors
- Reimsnider, S., Manfredsson, F. P., Muzyczka, N., & Mandel, R. J.
- Affiliations
- University of Florida Health (US)
- Corresponding author
- Mandel, Ronald J
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2007-08
- Volume
- 15
- Issue
- 8
- Pages
- 1504-1511
- ISSN
- 1525-0016 (ISSN-L)
- OpenAlex
- W1983678138
- Cited by
- 125
- References
- 42
- Citations by year
- 2026: 7 · 2025: 6 · 2024: 6 · 2023: 6 · 2022: 3 · 2021: 4 · 2020: 7 · 2019: 4 · 2018: 5 · 2017: 4 · 2016: 6 · 2015: 5 · 2014: 5 · 2013: 11 · 2012: 4
- FWCI
- 5.53
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- University of Florida
- National Institute of Neurological Disorders and Stroke (P01NS036302, P01NS36302)
- 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
- Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
- Keywords
- recombinant adeno-associated virus vectors, intrastriatal injection, striatal neurons, green fluorescent protein, neuroinflammation, gene therapy
- MeSH terms
- Adenoviridae, Animals, Cell Differentiation, DNA, Recombinant, Female, Genetic Vectors, Neuroglia, Time Factors, Transduction, Genetic, Gene Expression, Rats, Sprague-Dawley, Genes, Reporter, Transgenes, Rats
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