Off-target effects of VEGF-A regulation by shRNAs

The aim of this project was to investigate how genome-wide gene expression patterns change when the expression of VEGF-A is modulated using different lentivirally delivered shRNA molecules that are complementary to VEGF-A promoter region. To study these changes, 12 gene expression array experiments were conducted with mouse endothelial cell line (C166). The C166 cells were transduced (20% confluency, MOI 10) with lentiviral vectors that all express GFP marker gene and two of them express additional shRNA molecule that is complementary to different regions in the VEGF-A promoter and either up-regulate (VEGF-up) or downregulate (VEGF-down) VEGF-A expression. Fresh medium was changed 72h after transduction. Cells were collected at day 7.