Cited by 190Virus-based gene therapy researchRNA Interference and Gene DeliveryCRISPR and Genetic Engineering

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Large‐scale molecular characterization of adeno‐associated virus vector integration in mouse liver

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Nakai, Hiroyuki, Wu, Xiaolin, Fuess, Sally, Storm, Theresa A., Munroe, David, Montini, Eugenio, Burgess, Shawn M., Grompe, Markus, Kay, Mark A.·2005·Journal of Virology

DOI 10.1128/jvi.79.6.3606-3614.2005PMC PMC1075691

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Abstract

Recombinant adeno-associated virus (rAAV) vector holds promise for gene therapy. Despite a low frequency of chromosomal integration of vector genomes, recent studies have raised concerns about the risk of rAAV integration because integration occurs preferentially in genes and accompanies chromosomal deletions, which may lead to loss-of-function insertional mutagenesis. Here, by analyzing 347 rAAV integrations in mice, we elucidate novel features of rAAV integration: the presence of hot spots for integration and a strong preference for integrating near gene regulatory sequences. The most prominent hot spot was a harmless chromosomal niche in the rRNA gene repeats, whereas nearly half of the integrations landed near transcription start sites or CpG islands, suggesting the possibility of activating flanking cellular disease genes by vector integration, similar to retroviral gain-of-function insertional mutagenesis. Possible cancer-related genes were hit by rAAV integration at a frequency of 3.5%. In addition, the information about chromosomal changes at 218 integration sites and 602 breakpoints of vector genomes have provided a clue to how vector terminal repeats and host chromosomal DNA are joined in the integration process. Thus, the present study provides new insights into the risk of rAAV-mediated insertional mutagenesis and the mechanisms of rAAV integration.

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Authors: Nakai, H., Wu, X., Fuess, S., Storm, T. A., Munroe, D., Montini, E., Burgess, S. M., Grompe, M., & Kay, M. A.
Affiliations: Stanford Medicine (US)
National Institutes of Health (US)
Oregon Health & Science University (US)
Corresponding author: Nakai, Hiroyuki
ORCID: Nakai, Hiroyuki
Wu, Xiaolin
Storm, Theresa A.
Montini, Eugenio
Burgess, Shawn M.
Grompe, Markus
Kay, Mark A.
Venue: Journal of Virology
Publisher: American Society for Microbiology
Published: 2005-03-15
Volume: 79
Issue: 6
Pages: 3606-3614
ISSN: 0022-538X (ISSN-L)
1098-5514
DOI: 10.1128/jvi.79.6.3606-3614.2005
PMC: PMC1075691
OpenAlex: W2151713466
Cited by: 190
References: 33
Citations by year: 2026: 4 · 2025: 2 · 2024: 4 · 2023: 7 · 2022: 7 · 2021: 1 · 2020: 11 · 2019: 4 · 2018: 5 · 2017: 3 · 2016: 10 · 2015: 13 · 2014: 8 · 2013: 6 · 2012: 13
FWCI: 10.25
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Journal of Virology (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion)
PubMed (publishedVersion)
PubMed Central (publishedVersion)
Funders: U.S. Department of Health and Human Services
National Institutes of Health (n01co12400-79-0-22, N01-CO-12400)
National Heart, Lung, and Blood Institute (HL66948, U01HL066948)
National Cancer Institute (N01CO12400, N01-CO-012400)
National Institute of Diabetes and Digestive and Kidney Diseases (DK18252, DK068636, R21DK068636)
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 › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
Keywords: adeno-associated virus, insertional mutagenesis, liver, CpG islands, transcription start sites, cancer genes, chromosomal breakpoints, terminal repeats, gene therapy safety, mice
MeSH terms: Dependovirus, Animals, Genetic Vectors, Oncogenes, Regulatory Sequences, Nucleic Acid, Genetic Therapy, Mutagenesis, Insertional, Virus Integration, CpG Islands, Genes, rRNA, Hepatocytes, Transcription Initiation Site, Mice

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Large‐scale molecular characterization of adeno‐associated virus vector integration in mouse liver

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