SC35 CRISPR Activation Plasmid (h): sc-417676-ACT

Human SRSF2 (SC35) encodes a serine/arginine-rich splicing factor that binds pre-mRNA and coordinates splice-site selection, exon definition, and coupling of transcription with RNA processing. SC35 participates in spliceosome assembly and influences alternative splicing programs that shape cell-cycle progression, DNA damage responses, and differentiation by modulating isoform output across regulatory gene networks. Perturbation of SRSF2 activity can rewire transcriptome-wide splicing patterns, altering protein domain composition and signaling pathway connectivity. Dysregulated SRSF2 expression or function is associated with aberrant RNA processing signatures observed in multiple disease-relevant cellular states, making it a useful node for mechanistic studies of splicing-dependent phenotypes.

SC35 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SRSF2 expression without altering the underlying DNA sequence.

SC35 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SRSF2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the SRSF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SC35 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SRSF2 locus and enabling the study of SC35-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SC35 pathway restoration in tumor cells with silenced or reduced SRSF2 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.