Genetic Analysis of Anopheles mosquitoes

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DESCRIPTION (provided by applicant): Insect vector-borne diseases continue to burden a large fraction of the world's population. Understanding the basic biology of the insects responsible for transmitting the pathogens that cause these diseases has been and will continue to be essential for the development of effective strategies of control. Technological advances often provide novel opportunities to enlarge our understanding of basic biology. This project will result in the creation of enhancer- and gene-trap technologies for the human malaria mosquito Anopheles stephensi that will enable researchers to identify, isolate and analyze genes in new and powerful ways. The technology will be applied to the study of the salivary glands, an organ that plays a vital role in the transmission of pathogens such as Plasmodium. Preliminary results show that the transposable element piggyBac is highly active within the genome of Anopheles stephensi, making it a useful platform upon which to build these genetic technologies. Prototype enhancer- trap and gene-trap systems have been created and introduced into Anopheles stephensi. Preliminary screens to test the functionality of both systems were successfully completed. Both systems are active and effective at sensing enhancers and genes, respectively. A small diverse collection of enhancer-trap lines is described which provides a proof of principle. Based on prototype designs, existing Anopheles stephensi enhancer- and gene-trap technologies will be modified for increased efficiency and effectiveness. The transgenic lines necessary for conducting enhancer- and gene-trap screens, as well as, useful 'Gal4 driver lines' will become a community resources. Anopheles stephensi gene-trap technology will be used to identify and isolate genes specifically expressed in the salivary glands of larvae and adults under a variety of developmental and physiological conditions. This project will enable the importance of various salivary gland secretions in mosquito feeding and pathogen transmission to be understood, and to integrate knowledge concerning salivary gland structure and function. This new detailed understanding of this critical tissue in a major vector o human disease is expected to lead to the conception and implementation of new strategies for disrupting pathogen transmission.