4.3 Article

Generation of a new Gateway-compatible inducible lentiviral vector platform allowing easy derivation of co-transduced cells

Journal

BIOTECHNIQUES
Volume 60, Issue 5, Pages 252-259

Publisher

BIOTECHNIQUES OFFICE
DOI: 10.2144/000114417

Keywords

lentiviral vectors; co-inducible expression; Tet-On; Gateway

Funding

  1. VIB
  2. European grant: FP6 Epistem [LSHB-CT-2005-019067]
  3. European grant: FP7 TuMIC [2008-201662]
  4. European grant: Euregional PACT II
  5. Belgian grants: Interuniversity Attraction Poles [IAP 6/18, IAP7]
  6. Stichting tegen Kanker [2010-162]
  7. Flemish grants: FWO-Vlaanderen [1.5.058.12N, G.0544.11, G.0226.09, 1.5.169.08N, G.0A45.12N, G.0214.09N, G.0942.10N]
  8. Ghent University grants: MRP, GROUP-ID consortium, Concerted Research Actions [01G019A8-B8]
  9. Flemish Government [BOF09/01M00709]
  10. Instituut voor Innovate door Wetenschap en Technologie
  11. Fonds voor Wetenschappelijk Onderzoek-Vlaanderen

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In contrast to most common gene delivery techniques, lentiviral vectors allow targeting of almost any mammalian cell type, even non-dividing cells, and they stably integrate in the genome. Therefore, these vectors are a very powerful tool for biomedical research. Here we report the generation of a versatile new set of 22 lentiviral vectors with broad applicability in multiple research areas. In contrast to previous systems, our platform provides a choice between constitutive and/or conditional expression and six different C-terminal fusions. Furthermore, two compatible selection markers enable the easy derivation of stable cell lines co-expressing differently tagged transgenes in a constitutive or inducible manner. We show that all of the vector features are functional and that they contribute to transgene overexpression in proof-of-principle experiments.

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