The long-term goal of this research is to develop and apply molecular genetic techniques for the control of arthropod-borne virus (arbovirus) diseases. Conventional control measures, including pesticide application, environmental sanitation, and immunization have proven to be effective in short-term reduction of arbovirus diseases, but not in long-term reduction or eradication. Indeed, many arbovirus disease remain significant pathogens or are resurgent in previously controlled areas. In this proposal, novel molecular genetic strategies will be investigated for manipulation of the Aedes triseriatus genome to intracellularly immunize the mosquito to prevent to reduce LaCrosse (LAC) virus infection and transmission.
The specific aims are: 1. Intracellular immunization: The efficacy of developed antiviral constructs, which express specific virus proteins, virus antisense sequences, virus specific ribozymes, will determined using a Sindbis virus expression system. The Sindbis system permits expression of antiviral constructs in vitro in cell cultures and in vivo in mosquitoes. Constructs that interfere with virus infection, replication, and/or transmission will then be candidates for generation of transgenic mosquitoes. 2. The Aedes densonucleosis mosquitoes. Expression of transform mosquitoes by either genomic integration or as a stable episome will be tested. 4. An arbovirus refractory mosquito will be bioengineered by introducing promising antiviral constructs into the genome by transduction or other transformation procedures. In addition to providing important information about the potential of intracellular immunization for control of arbovirus (and other virus) disease, these studies will provide invaluable information about gene regulation and expression in vectors, and the potential for utilization of the new Aedes parvovirus as a tool for genetic manipulation of vectors. Considerable information should be forthcoming from these proposed studies which could provide novel strategies for control of these very important vecto-borne virus pathogens of humans and animals.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI025629-06A1
Application #
2063035
Study Section
Tropical Medicine and Parasitology Study Section (TMP)
Project Start
1988-07-01
Project End
1999-01-31
Budget Start
1994-05-01
Budget End
1995-01-31
Support Year
6
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Colorado State University-Fort Collins
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
112617480
City
Fort Collins
State
CO
Country
United States
Zip Code
80523
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Blair, C D; Adelman, Z N; Olson, K E (2000) Molecular strategies for interrupting arthropod-borne virus transmission by mosquitoes. Clin Microbiol Rev 13:651-61
Allen-Miura, T M; Afanasiev, B N; Olson, K E et al. (1999) Packaging of AeDNV-GFP transducing virus by expression of densovirus structural proteins from a sindbis virus expression system. Virology 257:54-61
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Kimmick, M W; Afanasiev, B N; Beaty, B J et al. (1998) Gene expression and regulation from the p7 promoter of Aedes densonucleosis virus. J Virol 72:4364-70
Kamrud, K I; Olson, K E; Higgs, S et al. (1998) Use of the Sindbis replicon system for expression of LaCrosse virus envelope proteins in mosquito cells. Arch Virol 143:1365-77
Seabaugh, R C; Olson, K E; Higgs, S et al. (1998) Development of a chimeric sindbis virus with enhanced per Os infection of Aedes aegypti. Virology 243:99-112
Kamrud, K I; Olson, K E; Higgs, S et al. (1997) Detection of expressed chloramphenicol acetyltransferase in the saliva of Culex pipiens mosquitoes. Insect Biochem Mol Biol 27:423-9
Powers, A M; Kamrud, K I; Olson, K E et al. (1996) Molecularly engineered resistance to California serogroup virus replication in mosquito cells and mosquitoes. Proc Natl Acad Sci U S A 93:4187-91
Corsini, J; Traul, D L; Wilcox, C L et al. (1996) Efficiency of transduction by recombinant Sindbis replicon virus varies among cell lines, including mosquito cells and rat sensory neurons. Biotechniques 21:492-7

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