Project description:Aedes aegypti is a vector of many infectious agents, including flaviviruses like Zika virus. We demonstrate that Nest1, a 34kDa mosquito salivary protein, facilitates Zika virus dissemination in human skin explants. Our aim was to analyze the effect of Nest1 at a transcriptomic level (RNAseq) in human skin explants, in the presence and absence of virus (ZIKV), at different timepoints (day 1, 2,3 ,and 4)
Project description:We report a global survey of viral small RNAs (vsmRNAs) from >200 Aedes aegypti samples to identify many mosquito viruses that actively infect this prominent arboviral vector. Ae. aegypti viruses in the Americas were abundant, with some displaying geographical boundaries. Viruses infecting Asian Ae. aegypti were similar to those in the Americas and revealed the first wild example of dengue vsmRNAs. African Ae. aegypti displayed vsmRNAs from viruses unique to these African strains. Academic lab colonies generally lacked viruses, yet two commercial strains were deeply infected by a tombus-like virus that is related to plant viruses. Comparing matched viral long RNAs to vsmRNAs revealed viral transcripts evading the mosquito RNA interference (RNAi) pathway. By infecting mosquito cells with Ae. aegypti homogenates, we generated stably infected cell lines which produced vsmRNAs that were comparable to native mosquito vsmRNA patterns. Lastly, we demonstrate that these stably infected mosquito cells producing vsmRNAs can exert gene silencing of reporters bearing viral sequence segments, providing a potential explanation for how Ae. aegypti can tolerate the persistence of viral infections. This vsmRNA genomics approach in Ae. aegypti can add to existing vector surveillance approaches by discovering new viruses that persist in mosquito populations.
Project description:Transcriptional profiling of mosquito head comparing control non-swarming mosquito with swarming mosquito. Screen the genes associated with swarming.
Project description:Anopheles gambiae mosquitoes are primary human malaria vectors, but we know very little about their mechanisms of transcriptional regulation. We profiled chromatin accessibility by ATAC-seq in laboratory-reared An. gambiae mosquitoes experimentally infected with the human malaria parasite Plasmodium falciparum. By integrating ATAC-seq, RNA-seq and ChIP-seq data we showed a positive correlation between accessibility at promoters and introns, gene expression and active histone marks. By comparing expression and chromatin structure patterns in different tissues, we were able to infer cis-regulatory elements controlling tissue specific gene expression and to predict the in vivo binding sites of relevant transcription factors. The ATAC-seq assay also allowed the precise mapping of active regulatory regions, including novel transcription start sites and enhancers that annotate to mosquito immune-response genes. This study is important not only for advancing our understanding of mechanisms of transcriptional regulation in the mosquito vector of human malaria, but also the information we produced is of great potential for developing new mosquito-control and anti-malaria strategies.
Project description:We have published a paper showing that infections using mosquito-transmitted parasites have a quite different effect on the host immune system (Spence et al, Nature, 2013). Here we are repeating the experiment but using a more virulent strain of the parasite: P. chabaudi CB, to study further how mosquito transmission regulates parasite virulence. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description:Aedes aegypti are vectors of several devastating arboviruses infecting hundreds of millions of people annually. Controlling mosquito populations by regulating their reproduction is important to minimize viral transmission in the absence of effective antiviral therapies or vaccines. Here, we demonstrate that leucine aminopeptidase1 (LAP1), screened from SWATH-MS-based proteomic data of female spermathecae, is a crucial determinant in mosquito population expansion. Mitochondrial defects and aberrant autophagy of sperm in LAP1 mutant males (LAP1-/-), prepared using CRISPR-Cas9 system, resulted in a reduction of reproduction in wild-type females that mated with them. Additionally, we found that the fitness of LAP1-/- males was strong enough to efficiently transmit genetic changes to mosquito populations through a low number of hatchable offspring, making it to be a promising opportunity to suppress mosquito populations using LAP1-/- males. Importantly, we provide a novel target gene for genetic drive, further amplifying the function of LAP1 in reducing mosquito populations.