Post-integration silencing of piggyBac transposable elements in Aedes aegypti.

Autor(es): Palavesam Azhahianambi; Esnault Caroline; O'Brochta David A

Resumo: The piggyBac transposon, originating in the genome of the Lepidoptera Trichoplusia ni, has a broad host range, making it useful for the development of a number of transposon-based functional genomic technologies including gene vectors, enhancer-, gene- and protein-traps. While capable of being used as a vector for the creation of transgenic insects and insect cell lines, piggyBac has very limited mobility once integrated into the genome of the yellow fever mosquito, Aedes aegypti. A transgenic Aedes aegypti cell line (AagPB8) was created containing three integrated piggyBac elements and the remobilization potential of the elements was tested. The integrated piggyBac elements in AagPB8 were transpositionally silent in the presence of functional transposase, which was shown to be capable of catalyzing the movement of plasmid-borne piggyBac elements in the same cells. The structural integrity of one of the integrated elements along with the quality of element-flanking DNA, which is known to influence transposition rates, were tested in D. melanogaster. The element was found to be structurally intact, capable of transposition and excision in the soma and germ-line of Drosophila melanogaster, and in a DNA sequence context highly conducive to element movement in Drosophila melanogaster. These data show that transpositional silencing of integrated piggyBac elements in the genome of Aedes aegypti appears to be a function of higher scale genome organization or perhaps epigenetic factors, and not due to structural defects or suboptimal integration sites.

Palavras-Chave: Transposable elements; Invertebrate genomics; Drosophila melanogaster; Polymerase chain reaction; Aedes aegypti; Silencer elements; Functional genomics; Plasmid construction

Imprenta: PloS One, v. 8, n. 7, e68454, 2013

Identificador do objeto digital: 10.1371/journal.pone.0068454.

Descritores: Aedes aegypti - Cell ; Aedes aegypti - DNA ; Aedes aegypti - Genome ; Aedes aegypti - Molecular Structure ; Aedes aegypti - Pathogenesis ; Aedes aegypti - virus

Data de publicação: 2013