RNA-seq of human blood samples from MiBLEND study on the effect of fruit and vegetable blends on gene expression

This study investigates the effects of fruit and vegetable (F&V) intake on biomarkers of chronic diseases in healthy human volunteers. The study uses a randomized crossover design with a total duration of 7 weeks. Each participant underwent a 2-week run-in period to standardize F&V intake at 50g per day, followed by two separate 2-week intervention phases, during which F&V intake was increased to 450g per day, with a 1-week washout period between interventions. Participants were randomized to receive two out of seven different nutrition interventions, each providing distinct combinations of F&V. Blood samples were collected at baseline and after each intervention phase. Whole blood was drawn into EDTA-coated tubes, and RNA was extracted using the Directzol RNA microprep kit (ZY-R2052, Zymo Research), following ribo-depletion of rRNA. RNA quality and concentration were checked using a Qubit Fluorometer and Bioanalyzer. Samples with RIN values greater than 7 were used for library preparation. Library preparation was performed using the NextFlex Rapid Directional RNA kit 2.0 with ribo-depletion, followed by sequencing on the Illumina platform (200-cycle S4 flow cells). Sequencing generated paired-end reads of 150bp from approximately 150 samples per flow cell. The resulting fastq files were processed, de-multiplexed, and reviewed for quality, and all samples from each participant were analyzed in batches. Data analysis focused on differential gene expression related to antioxidant capacity, inflammation, and DNA damage markers.