Tn5 as an insect gene vector

Insect Biochemistry and Molecular Biology
Kathryn H RowanDavid A O'Brochta

Abstract

The purpose of this study was to explore alternatives to insect-derived transposable elements as insect gene vectors with the intention of improving existing insect transgenesis methods. The mobility properties of the bacterial transposon, Tn5, were tested in mosquitoes using a transient transposable element mobility assay and by attempting to create transgenic insects. Tn5 synaptic complexes were assembled in vitro in the absence of Mg(2+) and co-injected with a target plasmid into developing yellow fever mosquito, Aedes aegypti, embryos. Target plasmids recovered from embryos a day later were screened for the presence of Tn5. Recombinants (transposition events) were found at a frequency of 1.2 x 10(-3). Some transposition events did not appear to be associated with canonical 9 bp direct duplications at the site of insertion and also were associated with either deletions or rearrangements. A Tn5 element containing the brain-specific transgene, 3 x P3DsRed, was assembled into synaptic complexes in vitro and injected into pre-blastoderm embryos of Ae. aegypti. Of the approximately 900 embryos surviving injection and developing into adults, two produced transgenic progeny. Both transgenic events involved the co-integrations of ap...Continue Reading

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Citations

Dec 14, 2011·Annual Review of Entomology·Malcolm J Fraser
Aug 27, 2005·Journal of Medical Entomology·Nagaraja Sethuraman, David A O'Brochta
Aug 1, 2006·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·K J MaragathavallyC J Coates
Dec 26, 2018·Parasites & Vectors·Irina Häcker, Marc F Schetelig

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