Construction of stable packaging cell lines for clinical lentiviral vector production

Sanber, Khaled S.; Knight, Sean B.; Stephen, Sam L.; Bailey, Ranbir; Escors, David; Minshull, Jeremy; Santilli, Giorgia; Thrasher, Adrian J.; Collins, Mary K.; Takeuchi, Yasuhiro

doi:10.1038/srep09021

Cited by 108Virus-based gene therapy researchViral Infectious Diseases and Gene Expression in InsectsRNA Interference and Gene Delivery

Construction of stable packaging cell lines for clinical lentiviral vector production

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Sanber, Khaled S., Knight, Sean B., Stephen, Sam L., Bailey, Ranbir, Escors, David, Minshull, Jeremy, Santilli, Giorgia, Thrasher, Adrian J. +2 more·2015·Scientific Reports

DOI 10.1038/srep09021PMC PMC4356972

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Abstract

Lentiviral vectors are useful experimental tools for stable gene delivery and have been used to treat human inherited genetic disorders and hematologic malignancies with promising results. Because some of the lentiviral vector components are cytotoxic, transient plasmid transfection has been used to produce the large batches needed for clinical trials. However, this method is costly, poorly reproducible and hard to scale up. Here we describe a general method for construction of stable packaging cell lines that continuously produce lentiviral vectors. This uses Cre recombinase-mediated cassette exchange to insert a codon-optimised HIV-1 Gag-Pol expression construct in a continuously expressed locus in 293FT cells. Subsequently Rev, envelope and vector genome expression cassettes are serially transfected. Vector titers in excess of 10 6 transducing units/ml can be harvested from the final producer clones, which can be increased to 10 8 TU/ml by concentration. This method will be of use to all basic and clinical investigators who wish to produce large batches of lentiviral vectors.

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A method for generating clinical-grade stable packaging cell lines that continuously produce lentiviral vectors was reported.
“Recently, a method for generating clinical-grade stable packaging cell lines that continuously produce lentiviral vectors was reported [65].”
From Clinical use of lentiviral vectors (2018) · cites this paper as [65]
The Abstract (page unavailable) reports a method for constructing stable packaging cell lines that continuously produce lentiviral vectors. But the Methods > Cell Culture section describes the derived lines as potentially for future clinical use; our copy does not establish that the lines generated were already clinical-grade. (LLM, conf=0.86)
Experimental — accuracy not yet measured
The reported method for generating stable packaging cell lines solved a key issue with lentiviral vector production found in previous studies, which was that plasmid transfection of the gag-pol cassette did not induce continuous high-level expression.
“This approach solved a key issue with lentiviral vector production found in previous studies, which was that plasmid transfection of the gag-pol cassette did not induce continuous high-level expression.”
From Clinical use of lentiviral vectors (2018) · cites this paper as [65]

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A New-Generation Stable Inducible Packaging Cell Line for Lentiviral Vectors
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Authors: Sanber, K. S., Knight, S. B., Stephen, S. L., Bailey, R., Escors, D., Minshull, J., Santilli, G., Thrasher, A. J., Collins, M. K., & Takeuchi, Y.
Affiliations: University College London (GB)
ATUM (United States) (US)
Corresponding author: Takeuchi, Yasuhiro
ORCID: Sanber, Khaled S.
Knight, Sean B.
Escors, David
Minshull, Jeremy
Santilli, Giorgia
Thrasher, Adrian J.
Takeuchi, Yasuhiro
Venue: Scientific Reports
Publisher: Springer Science and Business Media LLC
Published: 2015-03-12
Volume: 5
Issue: 1
Pages: 9021-9021
ISSN: 2045-2322 (ISSN-L)
DOI: 10.1038/srep09021
PMC: PMC4356972
OpenAlex: W2079000181
Cited by: 108
References: 48
Citations by year: 2026: 8 · 2025: 8 · 2024: 14 · 2023: 10 · 2022: 5 · 2021: 9 · 2020: 9 · 2019: 11 · 2018: 15 · 2017: 8 · 2016: 8 · 2015: 3
FWCI: 7.79
Type: journal article
Language: en
Open access: gold
doi.org
Licence: cc-by
creativecommons.org/licenses/by/4.0 (tdm)
creativecommons.org/licenses/by/4.0 (vor)
Locations: Scientific Reports (publishedVersion, cc-by)
Europe PMC (PubMed Central) (publishedVersion, cc-by)
PubMed Central (publishedVersion, cc-by)
PubMed (publishedVersion)
UCL Discovery (University College London) (submittedVersion)
Research Explorer (The University of Manchester) (submittedVersion)
Funders: Wellcome Trust
European Commission
Health and Safety Executive
Medical Research Council
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords: lentiviral vector production, vector titer, gene therapy vectors
MeSH terms: Genetic Vectors, Humans, Protein Precursors, Retroviridae, Viral Envelope Proteins, HIV-1, Gene Expression, Lentivirus, Virus Assembly, gag Gene Products, Human Immunodeficiency Virus, pol Gene Products, Human Immunodeficiency Virus, rev Gene Products, Human Immunodeficiency Virus, HEK293 Cells, Homologous Recombination

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