Study of the RNA splicing in METTL4 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 N6-methylation of Am30 in U2 snRNA on splicing rate and accuracy in HeLa S3 cells. High-throughput polyA+ RNA-seq was performed for METTL4 methyltransferase gene knockout cells. This methyltransferase is responsible for indicated modification in U2 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.