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Post-integration stabilization of a transposon vector by terminal sequence deletion in Drosophila melanogaster

Nature Biotechnology volume 22, pages 1150–1154 (2004)Cite this article

Abstract

Germline transformation systems for nearly 20 insect species have been derived from transposable elements, allowing the development of transgenic insects for basic and applied studies1,2,3. These systems use a defective nonautonomous vector that results in stable vector integrations after the disappearance of transiently provided transposase helper plasmid4, which is essential to maintain true breeding lines and consistent transgene expression that would otherwise be lost after vector remobilization. The risk of remobilization by an unintended transposase source has so far not been a concern for laboratory studies, but the prospective use of millions of transgenic insects in biocontrol programs will likely increase the risk, therefore making this a critical issue for the ecological safety of field release programs5,6. Here we describe an efficient method that deletes a terminal repeat sequence of a transposon vector after genomic integration. This procedure prevents transposase-mediated remobilization of the other terminal sequence and associated genes, ensuring their genomic stability.

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Figure 1: Methodology for achieving transgene stabilization using the pBac{L1-PUbDsRed1-L2-3xP3-ECFP-R1} vector.
Figure 2: PCR analysis of the vector sequences in lines F34 and F34-1M.

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Acknowledgements

Grateful appreciation is extended to Rod Nagoshi for discussions on the development of stabilization vectors and to the US Department of Agriculture–National Research Initiative Competitive Grants Program for support of this research.

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Authors and Affiliations

  1. US Department of Agriculture, Center for Medical, Agricultural, and Veterinary Entomology, Agricultural Research Service, 1700 SW 23rd Drive, Gainesville, 32608, Florida, USA

    Alfred M Handler & Grazyna J Zimowska

  2. Max-Planck-Institut für molekulare Genetik, Center for Cardiovascular Research, Hessische Strasse 3-4, Berlin, D-10115, Germany

    Carsten Horn

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  1. Alfred M Handler
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Correspondence to Alfred M Handler.

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Handler, A., Zimowska, G. & Horn, C. Post-integration stabilization of a transposon vector by terminal sequence deletion in Drosophila melanogaster. Nat Biotechnol 22, 1150–1154 (2004). https://doi.org/10.1038/nbt1002

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