Rapid Uncoating of Vector Genomes Is the Key toEfficient Liver Transduction with Pseudotyped Adeno-Associated VirusVectors
Open accessAbstract
Transduction of the liver with single-stranded adeno-associated virus serotype 2 (AAV2) vectors is inefficient; less than 10% of hepatocytes are permissive for stable transduction, and transgene expression is characterized by a lag phase of up to 6 weeks. AAV2-based vector genomes packaged inside AAV6 or AAV8 capsids can transduce the liver with higher efficiency, but the molecular mechanisms underlying this phenomenon have not been determined. We now show that the primary barrier to transduction of the liver with vectors based on AAV2 capsids is uncoating of vector genomes in the nucleus. The majority of AAV2 genomes persist as encapsidated single-stranded molecules within the nucleus for as long as 6 weeks after vector administration. Double-stranded vector genomes packaged inside AAV2 capsids are at least 50-fold more active than single-stranded counterparts, but these vectors also exhibit a lag phase before maximal gene expression. Vector genomes packaged inside AAV6 or AAV8 capsids do not persist as encapsidated molecules and are more biologically active than vector genomes packaged inside AAV2 capsids. Our data suggest that the rate of uncoating of vector genomes determines the ability of complementary plus and minus single-stranded genomes to anneal together and convert to stable, biologically active double-stranded molecular forms.
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Different kinetics for the two serotypes have been described previously with gene addition approaches wherein higher doses of AAV correlated with higher levels of gene expression.
“Different kinetics for the two serotypes have been described previously with gene addition approaches wherein higher doses of AAV correlated with higher levels of gene expression.36”
From Adeno‐associated virus gene repair corrects a mouse model of hereditary tyrosinemia in vivo†‡ (2010) · cites this paper as [36]
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- Authors
- Thomas, C. E., Storm, T. A., Huang, Z., & Kay, M. A.
- Affiliations
- Stanford Medicine (US)
- Corresponding author
- Kay, Mark A.
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 2004-03-15
- Volume
- 78
- Issue
- 6
- Pages
- 3110-3122
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC353747
- OpenAlex
- W2113444875
- Cited by
- 358
- References
- 4
- Citations by year
- 2026: 3 · 2025: 6 · 2024: 7 · 2023: 9 · 2022: 12 · 2021: 7 · 2020: 12 · 2019: 12 · 2018: 9 · 2017: 11 · 2016: 7 · 2015: 20 · 2014: 9 · 2013: 16 · 2012: 22
- FWCI
- 17.67
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Journal of Virology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Heart, Lung, and Blood Institute (HL66948, U01HL066948, HL64274, R01HL064274)
- 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 › Epidemiology › Hepatitis B Virus Studies
- Keywords
- adeno-associated virus vectors, AAV2, liver transduction, AAV6, AAV8, single-stranded DNA, double-stranded DNA, lag phase, hepatocyte transduction, pseudotyping
- MeSH terms
- Dependovirus, Animals, Capsid, Female, Genetic Vectors, Liver, Mice, Inbred BALB C, Mice, Inbred C57BL, Transduction, Genetic, Virion, Genome, Viral, Gene Transfer Techniques, Transgenes, Hepatocytes, Mice
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