SRp40 CRISPR Activation Plasmid (h): sc-401886-ACT

Human SRSF5 encodes the splicing factor SRp40, a serine/arginine-rich RNA-binding protein that regulates constitutive and alternative pre-mRNA splicing and influences exon definition through interactions with exonic splicing enhancers. SRp40 contributes to RNA processing networks that couple splicing with transcription and mRNA export, shaping isoform choice in pathways controlling cell cycle progression, apoptosis, and stress responses. Altered SRSF5 activity or expression can shift splice isoform balance and has been associated with dysregulated RNA metabolism observed across multiple disease contexts, including cancer-related signaling and proliferative phenotypes. As a node in SR protein–mediated spliceosome regulation, SRp40 is frequently studied for its impact on transcriptome remodeling and proteome diversity.

SRp40 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SRSF5 expression without altering the underlying DNA sequence.

SRp40 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SRSF5 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 SRSF5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SRp40 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SRSF5 locus and enabling the study of SRp40-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SRp40 pathway restoration in tumor cells with silenced or reduced SRSF5 expression.

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