Conditional expression in the malaria mosquito Anopheles stephensi with Tet-On and Tet-Off systems

Genetics
Gareth J LycettThanasis G Loukeris

Abstract

We report successful conditional gene expression in the malaria vector, Anopheles stephensi, on the basis of binary systems consisting of gene driver and responder transgenic lines generated by Minos-mediated germline transformation. An A. gambiae tissue-specific enhancer derived from a serpin (SRPN10) gene was utilized to control the temporal and spatial expression of doxycycline (dox)-sensitive transcriptional regulators in the driver lines. The "Tet-Off" driver utilized the tetracycline-controlled transcriptional activator (tTA) that is unable to bind and activate transcription from tetracycline operators (TetO) in the presence of dox; the "Tet-on" driver utilized the reverse tTA (rtTA) that, conversely, binds and activates TetO operators in the presence of dox. The responder lines carried insertions encompassing a LacZ reporter gene, cis-regulated by a TetO-P-element hybrid promoter. The progeny of crosses between driver and responder lines expressed beta-galactosidase under dual, tissue-specific and dox-mediated regulation. In adult rtTA/TetOPlacZ progeny, dox treatment rapidly induced beta-galactosidase activity throughout the midgut epithelium and especially in malaria parasite-invaded epithelial cells. Transactivator-de...Continue Reading

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Citations

Dec 12, 2012·Cell and Tissue Research·Salvador Hernández-MartínezMario H Rodríguez
Dec 11, 2008·Biotechnology Letters·Noriko KarasakiTakahiro Kusakabe
Apr 16, 2013·Proceedings of the National Academy of Sciences of the United States of America·Rebeca Carballar-LejarazúAnthony A James
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