ArticleHuman gene therapy2011
A versatile adeno-associated virus vector producer cell line method for scalable vector production of different serotypes.
Article in Human gene therapy, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- Gene modification: Exploring the potential in treating kidney diseases.Pharmacological research · 2026Review
- Survivin in cardiovascular diseases and its therapeutic potential.Vascular pharmacology · 2025Review
- Evaluation of a rapid multi-attribute combinatorial high-throughput UV-Vis/DLS/SLS analytical platform for rAAV quantification and characterization.Molecular therapy. Methods & clinical development · 2024Article
- Manufacturing Cell and Gene Therapies: Challenges in Clinical Translation.Annals of laboratory medicine · 2024Review
- Conditional splicing system for tight control of viral overlapping genes.Journal of virology · 2024Article
- Adeno-associated virus as a delivery vector for gene therapy of human diseases.Signal transduction and targeted therapy · 2024Review
- Development of Stable Packaging and Producer Cell Lines for the Production of AAV Vectors.Microorganisms · 2024Review
- Systematic comparison of rAAV vectors manufactured using large-scale suspension cultures of Sf9 and HEK293 cells.Molecular therapy : the journal of the American Society of Gene Therapy · 2024Article
- Inducible HEK293 AAV packaging cell lines expressing Rep proteins.Molecular therapy. Methods & clinical development · 2023Article
- Self-attenuating adenovirus enables production of recombinant adeno-associated virus for high manufacturing yield without contamination.Nature communications · 2022Article
- Generation and Characterization of a Skeletal Muscle Cell-Based Model Carrying One Single Gne Allele: Implications in Actin Dynamics.Molecular neurobiology · 2021Article
- AAV6 Vexosomes Mediate Robust Suicide Gene Delivery in a Murine Model of Hepatocellular Carcinoma.Molecular therapy. Methods & clinical development · 2020Article
- MicroRNA-based recombinant AAV vector assembly improves efficiency of suicide gene transfer in a murine model of lymphoma.Cancer medicine · 2020Article
- Light-sheet Fluorescence Microscopy for the Study of the Murine Heart.Journal of visualized experiments : JoVE · 2018Article
- Pharmacology of Recombinant Adeno-associated Virus Production.Molecular therapy. Methods & clinical development · 2018Review
- Nephron segment-specific gene expression using AAV vectors.Biochemical and biophysical research communications · 2018Article
- High-Density Recombinant Adeno-Associated Viral Particles are Competent Vectors for In Vivo Transduction.Human gene therapy · 2016Article
- The potential of adeno-associated viral vectors for gene delivery to muscle tissue.Expert opinion on drug delivery · 2014Review
- Production of adeno-associated virus (AAV) serotypes by transient transfection of HEK293 cell suspension cultures for gene delivery.Journal of virological methods · 2014Article
- Different protein composition and functional properties of adeno-associated virus-6 vector manufactured from the culture medium and cell lysates.Molecular therapy. Methods & clinical development · 2014Article
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5 authors.
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Abstract
Application of adeno-associated virus (AAV) vector in large animal studies and clinical trials often requires high-titer and high-potency vectors. A number of currently used vector production methods, based on either transient transfection or helper virus infection of cell lines, have their advantages and limitations. We previously developed a 293-cell-based producer cell line method for high-titer and high-potency AAV2 vectors. Similar to several other methods, however, it requires multiple cloning steps for the vector and packaging plasmids and a two-step transfection and selection for stable cell lines. Here we report a simplified method with several key improvements and advantages: (1) a one-step cloning of AAV vector cassette into the serotype-specific packaging plasmid; (2) a single plasmid transfection and selection for stable AAV vector producer cell lines; (3) high vector yields of different serotypes, e.g., AAV2, 8, and 9, upon infection with an E1A/E1B-deleted helper adenovirus; (4) efficient packaging of both single-stranded and double-stranded (self-complementary) AAV vectors; and (5) efficient packaging of large AAV cassettes such as a mini-dystrophin vector (5.0 kb). All cell lines were stable with growth rates identical to the parental 293 cells. The vector yields were consistent among serotypes, with 5 × 10(13) to 8 × 10(13) vector genome particles per Nunc cell factory (equivalent to 40 15-cm plates). The vectors showed high potency for in vitro and in vivo transduction. In conclusion, the simple and versatile AAV producer cell line method can be useful for large scale AAV vector production in preclinical and clinical studies.
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