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- Title
Identification of <italic>Aedes aegypti</italic> cis-regulatory elements that promote gene expression in olfactory receptor neurons of distantly related dipteran insects.
- Authors
Mysore, Keshava; Li, Ping; Duman-Scheel, Molly
- Abstract
Background: Sophisticated tools for manipulation of gene expression in select neurons, including neurons that regulate sexually dimorphic behaviors, are increasingly available for analysis of genetic model organisms. However, we lack comparable genetic tools for analysis of non-model organisms, including <italic>Aedes aegypti</italic>, a vector mosquito which displays sexually dimorphic behaviors that contribute to pathogen transmission. Formaldehyde-assisted isolation of regulatory elements followed by sequencing (FAIRE-seq) recently facilitated genome-wide discovery of putative <italic>A. aegypti</italic> cis-regulatory elements (CREs), many of which could be used to manipulate gene expression in mosquito neurons and other tissues. The goal of this investigation was to identify FAIRE DNA elements that promote gene expression in the olfactory system, a tissue of vector importance. Results: Eight <italic>A. aegypti</italic> CREs that promote gene expression in antennal olfactory receptor neurons (ORNs) were identified in a <italic>Drosophila melanogaster</italic> transgenic reporter screen. Four CREs identified in the screen were cloned upstream of <italic>GAL4</italic> in a transgenic construct that is compatible with transformation of a variety of insect species. These constructs, which contained FAIRE DNA elements associated with the <italic>A. aegypti odorant coreceptor</italic> (<italic>orco</italic>), <italic>odorant receptor 1</italic> (<italic>Or1</italic>), <italic>odorant receptor 8</italic> (<italic>Or8</italic>) and <italic>fruitless</italic> (<italic>fru</italic>) genes, were used for transformation of <italic>A. aegypti</italic>. Six <italic>A. aegypti</italic> strains, including strains displaying transgene expression in all ORNs, subsets of these neurons, or in a sex-specific fashion, were isolated. The CREs drove transgene expression in <italic>A. aegypti</italic> that corresponded to endogenous gene expression patterns of the <italic>orco</italic>, <italic>Or1</italic>, <italic>Or8</italic> and <italic>fru</italic> genes in the mosquito antenna. CRE activity in <italic>A. aegypti</italic> was found to be comparable to that observed in <italic>D. melanogaster</italic> reporter assays. Conclusions: These results provide further evidence that FAIRE-seq, which can be paired with <italic>D. melanogaster</italic> reporter screening to test FAIRE DNA element activity in select tissues, is a useful method for identification of mosquito cis-regulatory elements. These findings expand the genetic toolkit available for the study of <italic>Aedes</italic> neurobiology. Moreover, given that the CREs drive comparable olfactory neural expression in both <italic>A. aegypti</italic> and <italic>D. melanogaster</italic>, it is likely that they may function similarly in multiple dipteran insects, including other disease vector mosquito species.
- Subjects
MOSQUITOES; GENE expression; GENETIC models; NEURONS; NEUROBIOLOGY
- Publication
Parasites & Vectors, 2018, Vol 11, Issue 1, pN.PAG
- ISSN
1756-3305
- Publication type
Article
- DOI
10.1186/s13071-018-2982-6