Female mosquitoes require a blood meal for reproduction, and during blood feeding spread many of the devastating vector-borne diseases in humans. A deeper understanding of the molecular mechanisms linked to mosquito blood meal processes, reproductive events and anti-pathogen immunity is of particular importance for devising innovative vector control strategies. Based on strong preliminary data, we suggest that microRNAs (miRNA) play important and conserved roles in blood digestive functions of vector mosquitoes. We have developed antagomirs for depletion of miRNAs and their functional characterization in Aedes aegypti and Anopheles gambiae. We have identified two miRNAs with strong effects on blood digestion. Evaluation of spatiotemporal specificity of these miRNAs will be achieved by utilizing GAL4-UAS miRNA Tough Decoy or sponge loss-of-function transgenic mosquitoes. An important step in understanding miRNA function is determining their targets. A multi-algorithm computational target-prediction approach will be used for determination of the miRNA targets. Luciferase reporter constructs carrying 3 2UTRs of the candidate miRNA targets will then be transfected into mosquito cell lines, and their expression levels will be tested after ectopic co-transfection of a miRNA that is being studied. A phenotype rescue will be achieved by RNA interference knockdown of a target gene in the transgenic loss-of-function miRNA mosquitoes. As an alternative approach, we will identify miRNAs complementary to 32UTRs of genes encoding key enzymes involved in blood digestion and study their respective roles. This application presents a comprehensive approach for functional characterization of miRNAs in two of the most important mosquito vectors for human diseases.

Public Health Relevance

Female mosquitoes require a blood meal for reproduction, providing the underlying mechanism for the spread of many devastating vector-borne diseases in humans. A deeper understanding of the molecular mechanisms linked to mosquito blood meal processes, reproductive events and anti-pathogen immunity is of particular importance for devising innovative vector control strategies. This application presents a comprehensive approach to study the roles of small non-coding RNAs, microRNAs, in blood digestion of the two most important mosquito vectors of human diseases-Aedes aegypti and Anopheles gambiae.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI113729-01
Application #
8775119
Study Section
Special Emphasis Panel (ZRG1-IDM-M (03))
Program Officer
Costero, Adriana
Project Start
2014-06-15
Project End
2019-05-31
Budget Start
2014-06-15
Budget End
2015-05-31
Support Year
1
Fiscal Year
2014
Total Cost
$417,091
Indirect Cost
$137,626
Name
University of California Riverside
Department
Zoology
Type
Schools of Earth Sciences/Natur
DUNS #
627797426
City
Riverside
State
CA
Country
United States
Zip Code
92521
Zhao, Bo; Kokoza, Vladimir A; Saha, Tusar T et al. (2014) Regulation of the gut-specific carboxypeptidase: a study using the binary Gal4/UAS system in the mosquito Aedes aegypti. Insect Biochem Mol Biol 54:1-10
Liu, Shiping; Lucas, Keira J; Roy, Sourav et al. (2014) Mosquito-specific microRNA-1174 targets serine hydroxymethyltransferase to control key functions in the gut. Proc Natl Acad Sci U S A 111:14460-5