Design and Construction of Functional AAV Vectors
Abstract
Using the basic principles of molecular biology and laboratory techniques presented in this chapter, researchers should be able to create a wide variety of AAV vectors for both clinical and basic research applications. Basic vector design concepts are covered for both protein coding gene expression and small non-coding RNA gene expression cassettes. AAV plasmid vector backbones (available via AddGene) are described, along with critical sequence details for a variety of modular expression components that can be inserted as needed for specific applications. Protocols are provided for assembling the various DNA components into AAV vector plasmids in Escherichia coli, as well as for transferring these vector sequences into baculovirus genomes for large-scale production of AAV in the insect cell production system.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In recombinant AAV vectors, a desired transgene expression cassette is packaged into the capsids instead of the wild-type viral genome.
“In recombinant AAV vectors (rAAVs), a desired transgene expression cassette is packaged into the capsids instead of the wild-type (WT) viral genome [10].”
From Understanding Capsid Assembly and Genome Packaging for Adeno-Associated Viruses (2017) · cites this paper as [10]
To construct rAAVs, only the flanking cis-active ITRs have to be retained for packaging of the transgene cassette while the rep and cap genes can be supplied in trans.
“To construct rAAVs, only the flanking cis-active ITRs have to be retained for the packaging of the transgene cassette while the rep and cap genes can be supplied in trans [10].”
From Understanding Capsid Assembly and Genome Packaging for Adeno-Associated Viruses (2017) · cites this paper as [10]
Cited by
Sorted by citations
Adeno-Associated Virus (AAV) as a Vector for Gene Therapy
Naso, M. F., Tomkowicz, B., Perry, W. L., & Strohl, W. R. (2017) · BioDrugs · doi:10.1007/s40259-017-0234-5 · Cited by 1,362 · see citing works
Twenty-Five Years of Structural Parvovirology
Mietzsch, M., Pénzes, J. J., & Agbandje-McKenna, M. (2019) · Viruses · doi:10.3390/v11040362 · Cited by 197 · see citing works
Pharmacology of Recombinant Adeno-associated Virus Production
Penaud-Budloo, M., François, A., Clément, N., & Ayuso, E. (2018) · Molecular Therapy - Methods & Clinical Development · doi:10.1016/j.omtm.2018.01.002 · Cited by 177 · see citing works
Davidsson, M., Wang, G., Aldrin-Kirk, P., Cardoso, T., Nolbrant, S., Hartnor, M., Mudannayake, J., Parmar, M., & Björklund, T. (2019) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1910061116 · Cited by 142 · see citing works
Understanding Capsid Assembly and Genome Packaging for Adeno-Associated Viruses
Bennett, A., Mietzsch, M., & Agbandje-McKenna, M. (2017) · Future Virology · doi:10.2217/fvl-2017-0011 · Cited by 82 · see citing works
Manufacturing Clinical Grade Recombinant Adeno-Associated Virus Using Invertebrate Cell Lines
Kotin, R. M., & Snyder, R. O. (2017) · Human Gene Therapy · doi:10.1089/hum.2017.042 · Cited by 59 · see citing works
- Authors
- Gray, J. T., & Zolotukhin, S.
- Affiliations
- St. Jude Children's Research Hospital (US)
- University of Florida (US)
- Corresponding author
- Gray, John T.
- ORCID
- Gray, John T.
- Venue
- Methods in Molecular Biology
- Publisher
- Humana Press
- Published
- 2011-09-27
- Volume
- 807
- Pages
- 25-46
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- OpenAlex
- W2409154298
- Cited by
- 63
- References
- 0
- Citations by year
- 2026: 5 · 2025: 6 · 2024: 10 · 2023: 4 · 2022: 5 · 2021: 6 · 2020: 5 · 2019: 7 · 2018: 2 · 2017: 5 · 2016: 2 · 2015: 4 · 2014: 1 · 2013: 1
- FWCI
- 1.64
- Type
- book chapter
- Language
- en
- Open access
- closed
- Licence
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- 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 › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- adeno-associated virus vectors, gene expression cassette, protein-coding genes, baculovirus expression vector
- MeSH terms
- Dependovirus, Escherichia coli, Genetic Vectors, Plasmids, Transfection, Baculoviridae
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
