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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  Aedes aegypti , 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  A. aegypti  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  A. aegypti  CREs that promote gene expression in antennal olfactory receptor neurons (ORNs) were identified in a  Drosophila melanogaster  transgenic reporter screen. Four CREs identified in the screen were cloned upstream of  GAL4  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  A. aegypti odorant coreceptor  ( orco ),  odorant receptor 1  ( Or1 ),  odorant receptor 8  ( Or8 ) and  fruitless  ( fru ) genes, were used for transformation of  A. aegypti . Six  A. aegypti  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  A. aegypti  that corresponded to endogenous gene expression patterns of the  orco ,  Or1 ,  Or8  and  fru  genes in the mosquito antenna. CRE activity in  A. aegypti  was found to be comparable to that observed in  D. melanogaster  reporter assays.    Conclusions  These results provide further evidence that FAIRE-seq, which can be paired with  D. melanogaster  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  Aedes  neurobiology. Moreover, given that the CREs drive comparable olfactory neural expression in both  A. aegypti  and  D. melanogaster , it is likely that they may function similarly in multiple dipteran insects, including other disease vector mosquito species.

parasites and vectors

Identification of Aedes aegypti cis-regulatory elements that promote gene expression in olfactory receptor neurons of distantly related dipteran insects