, , 2015.
Mosquito immunobiology: the intersection of vector health and vector competence, Annu. Rev. Entomol, vol.63, pp.145-167, 2018. ,
The global distribution and burden of dengue, Nature, vol.496, pp.504-507, 2013. ,
Flavivirus susceptibility in Aedes aegypti, Arch. Med. Res, vol.33, pp.379-388, 2002. ,
Refining the global spatial limits of dengue virus transmission by evidence-based consensus, PLoS Negl. Trop. Dis, vol.6, p.1760, 2012. ,
Aedes aegypti uses RNA interference in defense against Sindbis virus infection, BMC Microbiol, vol.8, p.47, 2008. ,
A micrococcal nuclease homologue in RNAi effector complexes, Nature, vol.425, pp.411-414, 2003. ,
Cheating evolution: engineering gene drives to manipulate the fate of wild populations, Nat. Rev. Genet, vol.17, pp.146-159, 2016. ,
The dengue virus NS5 protein intrudes in the cellular spliceosome and modulates splicing, PLoS Pathog, vol.12, p.1005841, 2016. ,
, , 2007.
Expression analysis of Tudor-SN protein in mouse tissues, Tissue Cell, vol.45, pp.21-31, 2013. ,
Controlling vector-borne diseases by releasing modified mosquitoes, Nat. Rev. Microbiol, vol.16, pp.508-518, 2018. ,
, , 2016.
Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti, Proc. Natl. Acad. Sci. U S A, vol.103, pp.4198-4203, 2006. ,
Dynamic nucleolar targeting of dengue virus polymerase NS5 in response to extracellular pH, J. Virol, vol.90, pp.5797-5807, 2016. ,
The RNA binding protein Tudor-SN is essential for stress tolerance and stabilizes levels of stress-responsive mRNAs encoding secreted proteins in Arabidopsis, Plant Cell, vol.22, pp.1575-1591, 2010. ,
Human Tudor staphylococcal nuclease (Tudor-SN) protein modulates the kinetics of AGTR1-3'UTR granule formation, FEBS Lett, vol.588, pp.2154-2161, 2014. ,
, Tudor staphylococcal nuclease, 2012.
, J. Biol. Chem, vol.287, pp.18130-18141
aquasalis (Diptera: Culicidae), Mem. Inst. Oswaldo Cruz, vol.100, pp.545-547, 2005. ,
Emerging arboviruses: why today?, One Health, vol.4, pp.1-13, 2017. ,
Tudor staphylococcal nuclease links formation of stress granules and processing bodies with mRNA catabolism in Arabidopsis, Plant Cell, vol.27, pp.926-943, 2015. ,
The Tudor protein Veneno assembles the ping-pong amplification complex that produces viral piRNAs in Aedes mosquitoes, Nucleic Acids Res, vol.47, pp.2546-2559, 2019. ,
Aedes aegypti ML and Niemann-Pick type C family members are agonists of dengue virus infection, Dev. Comp. Immunol, vol.43, pp.1-9, 2014. ,
Dengue viruses cluster antigenically but not as discrete serotypes, Science, vol.349, pp.1338-1343, 2015. ,
RNA interference acts as a natural antiviral response to O'nyong-nyong virus (Alphavirus; Togaviridae) infection of Anopheles gambiae, Proc. Natl. Acad. Sci. U S A, vol.101, pp.17240-17245, 2004. ,
Tudor-SN interacts with piwi antagonistically in regulating spermatogenesis but synergistically in silencing transposons in Drosophila, PLoS Genet, vol.12, p.1005813, 2016. ,
New insights into nucleolar structure and function, 1000. ,
Tudor domain, Curr. Biol, vol.20, pp.666-667, 2010. ,
, , 2019.
, Mosquito antiviral defense mechanisms: a delicate balance between innate immunity and persistent viral infection, Parasit. Vectors, vol.12, p.165
Functional interaction between cellular p100 and the dengue virus 3' UTR, J. Gen. Virol, vol.92, pp.796-806, 2011. ,
Dengue virus infection of Aedes aegypti requires a putative cysteine rich venom protein, PLoS Pathog, vol.11, p.1005202, 2015. ,
Genetic dissection of Flaviviridae host factors through genome-scale CRISPR screens, Nature, vol.535, pp.159-163, 2016. ,
The epigenetic regulator g9a mediates tolerance to RNA virus infection in Drosophila, PLoS Pathog, vol.11, p.1004692, 2015. ,
The heat shock response restricts virus infection in Drosophila, Sci. Rep, vol.5, p.12758, 2015. ,
Beyond RNAi: antiviral defense strategies in Drosophila and mosquito, J. Insect Physiol, vol.59, pp.159-170, 2013. ,
The current and future global distribution and population at risk of dengue, Nat. Microbiol, vol.4, pp.1508-1515, 2019. ,
PIWIs go viral: arbovirus-derived piRNAs in vector mosquitoes, PLoS Pathog, vol.12, p.1006017, 2016. ,
Stress granules and virus replication, Future Virol, vol.6, pp.1329-1338, 2011. ,
Bugs are not to Be silenced: small RNA pathways and antiviral responses in insects, Annu. Rev. Virol, vol.3, pp.573-589, 2016. ,
URL : https://hal.archives-ouvertes.fr/pasteur-01957180
Alphavirus-derived small RNAs modulate pathogenesis in disease vector mosquitoes, Proc. Natl. Acad. Sci. U S A, vol.105, 2008. ,
Control of dengue virus in the midgut of Aedes aegypti by ectopic expression of the dsRNA-binding protein Loqs2, Nat. Microbiol, vol.3, pp.1385-1393, 2018. ,
Development of a TaqManâ RT-PCR assay without RNA extraction step for the detection and quantification of African Chikungunya viruses, J. Virol. Methods, vol.124, pp.65-71, 2005. ,
STATs as critical mediators of signal transduction and transcription: lessons learned from STAT5, Cytokine Growth Factor Rev, vol.15, pp.435-455, 2004. ,
Dengue virus replicates and accumulates in Aedes aegypti salivary glands, Virology, vol.507, pp.75-81, 2017. ,
URL : https://hal.archives-ouvertes.fr/pasteur-01976225
Individual co-variation between viral RNA load and gene expression reveals novel host factors during early dengue virus infection of the Aedes aegypti midgut, PLoS Negl. Trop. Dis, vol.11, p.6152, 2017. ,
URL : https://hal.archives-ouvertes.fr/pasteur-01953185
Dengue virus type 2: replication and tropisms in orally infected Aedes aegypti mosquitoes, BMC Microbiol, vol.7, p.9, 2007. ,
Viruses and the nucleolus: the fatal attraction, Biochim. Biophys. Acta, vol.1842, pp.840-847, 2014. ,
URL : https://hal.archives-ouvertes.fr/hal-00965059
, , 2009.
, PLoS Pathog, vol.5, p.1000299
Identification of Zika virus and dengue virus dependency factors using functional genomics, Cell Rep, vol.16, pp.232-246, 2016. ,
Discovery of insect and human dengue virus host factors, Nature, vol.458, pp.1047-1050, 2009. ,
Expanding the canon: non-classical mosquito genes at the interface of arboviral infection, Insect Biochem, 2019. ,
, Mol. Biol, vol.109, pp.72-80
Diverse host and restriction factors regulate mosquito-pathogen interactions, Trends Parasitol, vol.34, pp.603-616, 2018. ,
How does the royal family of Tudor rule the PIWIinteracting RNA pathway?, Genes Dev, vol.24, pp.636-646, 2010. ,
Nucleolus: the fascinating nuclear body, Histochem. Cell Biol, vol.129, pp.13-31, 2008. ,
URL : https://hal.archives-ouvertes.fr/hal-00195457
An evolutionary conserved function of the JAK-STAT pathway in anti-dengue defense, Proc. Natl. Acad. Sci. U S A, vol.106, pp.17841-17846, 2009. ,
Identification of viral suppressors of RNAi by a reporter assay in Drosophila S2 cell culture, Methods Mol. Biol, vol.721, pp.201-213, 2011. ,
Urbanization and geographic expansion of zoonotic arboviral diseases: mechanisms and potential strategies for prevention, Trends Microbiol, vol.21, pp.360-363, 2013. ,
Doublestranded RNA is produced by positive-strand RNA viruses and DNA viruses but not in detectable amounts by negative-strand RNA viruses, J. Virol, vol.80, pp.5059-5064, 2006. ,
Aedes aegypti control through modernized, integrated vector management, PLoS Curr, vol.9, 2017. ,
and 500 ng FLuc-specific and 1 µg GFP-, TSN-or Ago2-specific dsRNA. After incubation for 3 days at 28°C, mosquitoes were homogenized in passive lysis buffer (Promega) using the Precellys 24 grinder (Bertin Technologies) for 30 sec at 6,000 rpm. Samples were transferred to a 96-well plate and centrifugated for 5 min at 12000 x g. Fifty microliters of supernatant were transferred to a new plate, and 50 µL LARII reagent added for the first FLuc measurement. Next, 50 µL Stop&Glow reagent was added before the second measurement of RLuc, Nature, vol.535, pp.164-168, 2007. ,
DENV-1-infected Aag2 cells were fixed on coverslips using 4% paraformaldehyde (Sigma-Aldrich) for 30 min at room temperature (20-25°C) ,
, PBS, 0.1% Triton-X100, cells were incubated with mouse anti-dsRNA ?K1 (English & Scientific consulting) and rabbit anti-TSN antibodies diluted 1:500 in 1X PBS, 0.1% Triton-X100, p.2
, Normal Goat Serum for 1 hour at room temperature. Subsequently, cells were washed tree times with 1X PBS with 0.1% Triton X-100 and incubated with goat anti-mouse AlexaFluor 594 and goat anti-rabbit Alexa Fluor 488 diluted 1:1.000 in 1X PBS, 0.1% Triton-X100, 2% Normal Goat Serum (Life technologies)
, Triton X-100, cover slips were mounted on a glass slide in ~10 µL Prolong Gold anti-fade medium containing DAPI (Thermo Fisher) and imaged with a confocal microscope
, Infection with DENV-1 was performed 24 hours after the last transfection. Cells were incubated for 1 hour in L-15 infection medium containing 2% FBS and DENV-1 at a multiplicity of infection of 1. After removal of the infectious inoculum, cells were refreshed with fully supplemented with L-15 medium and incubated at 28°C. Western blotting Aag2 cells were harvested, Aag2 cells were transfected in 24-well plates with 500 ng of dsRNA using Lipofectamine LTX (Invitrogen) along with Plus reagent according to the manufacturer's instructions
HRP-conjugated polyclonal goat anti-rabbit IgG (GE Healthcare) diluted 1:5.000 in 1X PBS, 0.1% Triton-X100 for 1 hour at room temperature. Next, the blot was incubated with a primary anti-?-actin murine antibody (Sigma-Aldrich, clone AC-74) at 1:6.000 dilution in blocking buffer, and an HRP-conjugated polyclonal goat anti-mouse IgG (GE Healthcare) diluted at 1:5.000 in 1X PBS, 0.1% Triton-X100 was used as a secondary antibody. Bound antibodies were revealed by chemiluminescence with the SuperSignal West Pico Chemiluminescent Substrate, 1X PBS). After 3 washes with 1X PBS, the blot was incubated with a secondary antibody, 2017. ,
, Small RNA libraries
, Total RNA from pools of 2 (dsLuc) or 3 (dsTSN) whole mosquitoes was isolated with TRIzol (Invitrogen), p.15
, acrylamide/bisacrylamide (37.5:1), 7 M urea gel as described previously
Libraries were diluted to 4 nM and sequenced using an Illumina NextSeq 500 High Output kit v2 (75 cycles) on an Illumina NextSeq 500 platform. Sequence of fastq files was assessed using graphs generated by 'FastQC, ) on the fastq files created by 'cutadapt, 2009. ,
2009) to produce 'bam' indexed files. To analyze these 'bam' files, different kind of graphs were generated using home-made R scripts with several Bioconductor libraries such as 'Rsamtools' or 'Shortreads ,
Additionally, statistical significance tests as implanted in GraphPad Prism version 7. P values below 0, Statistical analysis methods have been described previously, p.5, 2017. ,
Validation of novel promoter sequences derived from two endogenous ubiquitin genes in transgenic Aedes aegypti, Insect Mol Biol, vol.19, pp.441-449, 2010. ,
First evidence of simultaneous circulation of three different dengue virus serotypes in Africa, PLoS ONE, vol.8, p.78030, 2013. ,
Alterations in the Aedes aegypti transcriptome during infection with West Nile, dengue and yellow fever viruses, PLoS Pathog, vol.7, p.1002189, 2011. ,
Quantifications of western blots with Image, 2017. ,
Genetic mapping of specific interactions between Aedes aegypti mosquitoes and dengue viruses, PLoS Genet, vol.9, 2013. ,
URL : https://hal.archives-ouvertes.fr/pasteur-00854586
Excretion of dengue virus RNA by Aedes aegypti allows non-destructive monitoring of viral dissemination in individual mosquitoes, Sci Rep, vol.6, p.24885, 2016. ,
URL : https://hal.archives-ouvertes.fr/pasteur-01316118
Viral small RNA cloning and sequencing, Methods Mol Biol, vol.721, pp.107-122, 2011. ,
URL : https://hal.archives-ouvertes.fr/pasteur-02089924
Virus-derived DNA drives mosquito vector tolerance to arboviral infection, Nat Commun, vol.7, p.12410, 2016. ,
URL : https://hal.archives-ouvertes.fr/pasteur-01377742
Regulated expression of microinjected DNA in adult Aedes aegypti mosquitoes, Insect Mol Biol, vol.16, pp.83-92, 2007. ,
Ultrafast and memory-efficient alignment of short DNA sequences to the human genome, Genome Biol, vol.10, p.25, 2009. ,
The Sequence Alignment/Map format and SAMtools, Bioinformatics, vol.25, pp.2078-2079, 2009. ,
Cutadapt removes adapter sequences from high-throughput sequencing reads, EMBnet Journal, vol.17, pp.10-12, 2011. ,
The epigenetic regulator g9a mediates tolerance to RNA virus infection in Drosophila, PLoS Pathog, vol.11, p.1004692, 2015. ,
The heat shock response restricts virus infection in Drosophila, Sci Rep, vol.5, 2015. ,
Individual co-variation between viral RNA load and gene expression reveals novel host factors during early dengue virus infection of the Aedes aegypti midgut, PLoS Negl Trop Dis, vol.11, p.6152, 2017. ,
URL : https://hal.archives-ouvertes.fr/pasteur-01953185
Whole-Genome Sequencing Analysis from the Chikungunya Virus Caribbean Outbreak Reveals Novel Evolutionary Genomic Elements, PLoS Negl Trop Dis, vol.10, p.4402, 2016. ,
URL : https://hal.archives-ouvertes.fr/pasteur-01634324
Identification of viral suppressors of RNAi by a reporter assay in Drosophila S2 cell culture, Methods Mol Biol, vol.721, pp.201-213, 2011. ,
Aedes aegypti Piwi4 Is a Noncanonical PIWI Protein, 2017. ,
URL : https://hal.archives-ouvertes.fr/pasteur-01573771