Modulating Splicing Factors by siRNA and Overexpression

The siRNA oligos targeting SRSF1, SRSF2, SRSF3, SRSF5, SRSF6, SRSF7, SRSF9, SRSF10, hnRNPM, hnRNPK, and hnRNPH1 were synthesized by Gene Pharma Company, the negative control si-NC was included. These siRNAs were transfected into cells with X-tremeGENE siRNA Transfection Reagent (ROCHE). The sequences were listed in Table S3.
The shRNAs specifically targeting SRSF2 (sh-SRSF2), SLMAP-Long isoform (sh-SLMAP-L), SLMAP-Short isoform (sh-SLMAP-S), CETN3-Long isoform (sh-CETN3-L), CETN3-Short isoform (sh-CETN3-S), and negative control (sh-Luci) were independently expressed in the knockdown lentiviruses vector PLKO.1-puro. The designed shRNAs were synthesized by Sangon Biotech. The shRNA sequences were listed in Table S4.
The wildtype SRSF2 (SRSF2-WT), SRSF2 domain deletion mutants (SRSF2-△RRM and SRSF2-△RS), the wildtype SRSF1 (SRSF1-WT), and wildtype hnRNPM (hnRNPM-WT) were constructed into pcDNA3.0 vector containing HA tag. Plasmids expressing SRSF2, SLMAP-L, SLMAP-S, CETN3-L, and CETN3-S transcript were constructed into PCDH-puro vector independently and synthesized by Sangon Biotech.