Study of the RNA splicing in THUMPD2 knockout HeLa S3 cells

Splicing is one of the most important stages in the maturation of the majority of RNA in eukaryotic cells. In this work, we studied the effect of m2G72 modification in U6 snRNA on splicing rate and accuracy in HeLa S3 cells. High-throughput polyA+ RNA-seq was performed for THUMPD2 methyltransferase gene knockout cells. This methyltransferase catalyzes the indicated N2-methylation of U6 snRNA. To study the splicing kinetics knockout cells were incubated for 10 and 20 minutes with 5-ethynyl uridine (EU), followed by nascent RNA purification and high throughput sequencing. Each experiment was done in 3 biological replicates.