Sleeping beauty transposition from nonintegrating lentivirus

Conrad A. Vink, H. Bobby Gaspar, Richard Gabriel, Manfred Schmidt, R S Mc Ivor, Adrian J. Thrasher, Waseem Qasim

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

Lentiviral vectors enter cells with high efficiency and deliver stable transduction through integration into host chromosomes, but their preference for integration within actively transcribing genes means that insertional mutagenesis following disruption of host proto-oncogenes is a recognized concern. We have addressed this problem by combining the efficient cell and nuclear entry properties of HIV-1-derived lentiviral vectors with the integration profile benefits of Sleeping Beauty (SB) transposase. Importantly, this integration enzyme does not exhibit a preference for integration within active genes. We generated integrase-deficient lentiviral vectors (IDLVs) to carry SB transposon and transposase expression cassettes. IDLVs were able to deliver transient transposase expression to target cells, and episomal lentiviral DNA was found to be a suitable substrate for integration via the SB pathway. The hybrid vector system allows genomic integration of a minimal promoter-transgene cassette flanked by short SB inverted repeats (IRs) but devoid of HIV-1 long terminal repeats (LTRs) or other virus-derived sequences. Importantly, integration site analysis revealed redirection toward a profile mimicking SB-plasmid integration and away from integration within transcriptionally active genes favored by integrase-proficient lentiviral vectors (ILVs).

Original languageEnglish (US)
Pages (from-to)1197-1204
Number of pages8
JournalMolecular Therapy
Volume17
Issue number7
DOIs
StatePublished - May 6 2009

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Beauty
Lentivirus
Transposases
Integrases
HIV-1
HIV Long Terminal Repeat
Genes
Proto-Oncogenes
Insertional Mutagenesis
Transgenes
Plasmids
Chromosomes
Viruses
DNA
Enzymes

Cite this

Vink, C. A., Gaspar, H. B., Gabriel, R., Schmidt, M., Mc Ivor, R. S., Thrasher, A. J., & Qasim, W. (2009). Sleeping beauty transposition from nonintegrating lentivirus. Molecular Therapy, 17(7), 1197-1204. https://doi.org/10.1038/mt.2009.94

Sleeping beauty transposition from nonintegrating lentivirus. / Vink, Conrad A.; Gaspar, H. Bobby; Gabriel, Richard; Schmidt, Manfred; Mc Ivor, R S; Thrasher, Adrian J.; Qasim, Waseem.

In: Molecular Therapy, Vol. 17, No. 7, 06.05.2009, p. 1197-1204.

Research output: Contribution to journalArticle

Vink, CA, Gaspar, HB, Gabriel, R, Schmidt, M, Mc Ivor, RS, Thrasher, AJ & Qasim, W 2009, 'Sleeping beauty transposition from nonintegrating lentivirus', Molecular Therapy, vol. 17, no. 7, pp. 1197-1204. https://doi.org/10.1038/mt.2009.94
Vink CA, Gaspar HB, Gabriel R, Schmidt M, Mc Ivor RS, Thrasher AJ et al. Sleeping beauty transposition from nonintegrating lentivirus. Molecular Therapy. 2009 May 6;17(7):1197-1204. https://doi.org/10.1038/mt.2009.94
Vink, Conrad A. ; Gaspar, H. Bobby ; Gabriel, Richard ; Schmidt, Manfred ; Mc Ivor, R S ; Thrasher, Adrian J. ; Qasim, Waseem. / Sleeping beauty transposition from nonintegrating lentivirus. In: Molecular Therapy. 2009 ; Vol. 17, No. 7. pp. 1197-1204.
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