Regulation of Splicing by Clinical Drugs. Homo sapiens

Alternative splicing analysis after treatment with three clinically aproved drugs Bioactive compounds have been invaluable for dissecting the mechanisms, regulation and functions of cellular processes. However, very few such reagents have been described for pre-mRNA splicing. To facilitate their systematic discovery, we developed a high throughput cell-based assay that measures pre-mRNA splicing utilizing a quantitative reporter system with advantageous features. The reporter, consisting of a destabilized, intron-containing luciferase expressed from a short-lived mRNA, allows rapid screens (23,000 screened), all pharmacologically active: clotrimazole, flunarizine and chlorhexidine. Interestingly, none was a general splicing inhibitor. Rather, each caused distinct splicing changes of numerous genes. Overall design: Treated HeLa cells with clotrimazole, flunarizine, chlorhexidine or DMSO for 6 hours followed by analysis of total RNA on human exon 1.0 ST platform. Three biological replicates were used for each treatment.