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First published online September 23, 2003

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The Journal of Experimental Biology 206, 3835-3843 (2003)
doi: 10.1242/jeb.00640

Review Article

Transgenesis and reverse genetics of mosquito innate immunity

Sang Woon Shin, Vladimir A. Kokoza and Alexander S. Raikhel^*

Department of Entomology, University of California, Riverside, California 92521, USA

^* Author for correspondence (e-mail: alexander.raikhel{at}ucr.edu)

Accepted 18 July 2003

In recent years, mosquito molecular biology has been a scene of astounding achievements, namely the development of genetic transformation, characterization of inducible tissue-specific promoters, and acquirement of mosquito genome sequences. However, the lack of a complete genetic tool box for mosquitoes remains a serious obstacle in our ability to study essential mosquito-specific mechanisms. Unlike Drosophila, very few null mutations for mosquito genes exist. The development of reverse-genetic analyses based on RNAi and transgenic techniques will help to compensate for these deficiencies and aid in identification of critical genes in important regulatory pathways. The study of mosquito innate immunity is one example and described here. In this study, we combine mosquito transgenesis with reverse genetics. The advantage of transgenesis is the ability to establish genetically stable, dominant-negative and overexpression phenotypes. Using the blood-meal-activated vitellogenin gene (Vg) promoter, we have generated transgenic mosquitoes with blood-meal-activated, overexpressed antimicrobial peptides, Defensin A and Cecropin A. Moreover, we have recently generated a transgenic dominant-negative Relish mosquito strain, which after taking a blood meal, becomes immune-deficient to infection by Gram-negative bacteria. The latter accomplishment has opened the door to a reverse-genetic approach in mosquitoes based on transgenesis.

Key words: transgenesis, reverse genetics, mosquito, immunity, defensin, cecropin, Relish, Dorsal, dsDNA, RNAi, Aedes aegypti, vitellogenin gene, transposable element

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