help button home button Genetics AJP: Regulatory Phys
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Genetics, Vol. 167, 1781-1790, August 2004, Copyright © 2004
doi:10.1534/genetics.104.028175

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lycett, G. J.
Right arrow Articles by Loukeris, T. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lycett, G. J.
Right arrow Articles by Loukeris, T. G.

Conditional Expression in the Malaria Mosquito Anopheles stephensi With Tet-On and Tet-Off Systems

Gareth J. Lycett*, Fotis C. Kafatos* and Thanasis G. Loukeris*,{dagger},1

* European Molecular Biology Laboratory, Heidelberg D-69117, Germany
{dagger} Institute of Molecular Biology and Biotechnology, FORTH, GR71110 Heraklion, Crete, Greece

1 Corresponding author: Institute of Molecular Biology and Biotechnology, FORTH, Vassilika Vouton, P.O. Box 1527, GR 711 10 Heraklion, Crete, Greece.
E-mail: loukeris{at}imbb.forth.gr

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 ß-galactosidase under dual, tissue-specific and dox-mediated regulation. In adult rtTA/TetOPlacZ progeny, dox treatment rapidly induced ß-galactosidase activity throughout the midgut epithelium and especially in malaria parasite-invaded epithelial cells. Transactivator-dependent, dox-mediated regulation was observed in hemocytes and pericardial cells using both systems. Conditional tissue-specific regulation is a powerful tool for analyzing gene function in mosquitoes and potentially for development of strategies to control disease transmission.






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2004 by the Genetics Society of America.