Introduction: The A. aegypti larvae are constantly exposed to environmental agents that can increase the production of reactive oxygen species (ROS). The excess of ROS can cause oxidative stress, damaging proteins, membranes and DNA. The DNA damage response (DDR) is responsible for identifying and repair lesions in DNA structure, ensuring genomic integrity. The study of DDR in A. aegypti may reveal essential steps for survival, reproduction or other biological process that can be exploited for insecticide development or as a blockage points to interfere in disease transmission. Objetive: Study DDR in A. aegypti larvae treated with paraquat through RNAseq. Materials and methods: A. aegypti larvae were treated with 0,5 mM of paraquat during 6h, 12h and 24h, and the RNAs were extracted with TRIzol. The RNAs were sent for cDNA library preparation and Illumina sequencing. Transcriptome analyses used Fastqc to measure sequencing quality and Cutadapt to remove adapters and sequences with poor quality. Reads were aligned against gene coding sequences (CDS) with Bowtie2 and Salmon was used for quantification. DESeq2 was used for identification of differentially expressed genes and Expander for generate expression profiles. Results: Sequencing (Single-end, 50bp) generated libraries (4 conditions in triplicate) with general quality higher than 30. We identified 1848 genes differentially expressed, that were grouped in 5 clusters. Key DDR genes such as PARP, XRCC1, XPC, XPB, HR23B, KU80, LIG4 and RAD50 were differentially expressed, highlighting the participation of base and nucleotide excision repair pathways, and the double strand break repair pathways, in the repair of damages caused by paraquat.

Study of DNA damage repair in Aedes aegypti larvae in response to oxidative stress

2019 Spring National Meeting

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