Project description:Chikungunya virus (CHIKV) is a single-stranded positive RNA virus that belongs to the genus Alphavirus and is transmitted to humans by infected Aedes aegypti and Aedes albopictus bites. In humans, CHIKV can cause painful symptoms during acute and chronic stages of infection. However, the virus-vector interaction has characteristics that allow a persistent infection, not disturbing the mosquito’s fitness. Here, we aimed to clarify aspects of CHIKV infection in Ae. aegypti Aag-2 cells through label-free quantitative proteomic analysis and transmission electron microscopy (TEM). We used MOI 0.1 to infect Aag-2 cells in biological triplicates over 48 h. TEM images show a high load of intracellular viral cargo at 48 hpi, as well as an elongated unusual mitochondria morphology that might indicate a mitochondrial imbalance. Moreover, a total of 196 Ae. aegypti protein groups were up or downregulated upon infection, related to protein synthesis, energy metabolism, signaling pathways and apoptosis. These regulated Aag-2 proteins might have roles in antiviral and/or in pro-viral mechanisms during CHIKV infection, to support the balance between viral propagation and the survival of host cell, leading to the persistent infection.
Project description:The impact of global climate change on the transmission dynamics of infectious diseases is the subject of extensive debate. The transmission of mosquito-borne viral diseases is particularly complex, with climatic variables directly affecting many parameters associated with the prevalence of disease vectors. While evidence shows that warmer temperatures often decrease the extrinsic incubation period of an arthropod-borne virus (arbovirus), exposure to cooler temperatures often predisposes disease vector mosquitoes to higher infection rates. RNA interference pathways are essential to antiviral immunity in the mosquito; however, few experiments have explored the effects of temperature on the RNAi machinery. Total small RNAs (miRNAs, siRNAs, piRNAs, etc.) were isolated and sequenced from the heads of sensor strain Aedes aegypti mosquitoes, or from the whole bodies of CHIKV-infected Aedes albopictus mosquitoes 8 hours post infection. Mosquitoes were grown at 18C or 28C in replicates of 1 (Ae. aegypti) or 3 (Ae. albopictus).
Project description:To determine codon optimality in Aedes Albopictus C6/36 cells, we blocked transcription using three independent transcription inhibitors (5,6-Dichlorobenzimidazole 1-β-D-ribofuranoside (DRB), Flavopiridol and Triptolide) and measured the RNA level at 6 hours post treatment using RNA-seq.
Project description:Ribosome profiling (Ribo-Seq) (maps positions of translating ribosomes on the transcriptome) and RNA-Seq (quantifies the transcriptome) analysis of African green monkey (Vero E6) cells and Aedes albopictus (C6/36) cells infected with Zika Virus (ZIKV) strain PE243. Cells were harvested at 24 h post infection (p.i.) and Ribo-Seq and RNA-Seq libraries were prepared and deep sequenced.