Dimethyl sulfoxide (DSMO) is a simple molecule widely used because of its great solvating ability. Beyond its physico-chemical properties, it is also biologically active, including on fungal species. Aspergillus flavus is a saprophytic and famous pathogenic fungus able to produce Aflatoxin B1 (AFB1), a potent carcinogenic mycotoxin which may contaminate many food crops. The aim of this study was to characterize the effect of DMSO on A. flavus transcriptome profile using high-throughput RNA-sequencing assay. Overall design: Six replicated fungal mycelium cultures were grown in three conditions (CTRL, DMSO Dose 1, DMSO Dose 2) to generate 18 samples in total which were extracted and processed to perform a RNAseq study.