SRp20 CRISPR Activation Plasmid (h): sc-401265-ACT

SRSF3 (SRp20) is a serine/arginine-rich splicing factor that regulates constitutive and alternative pre-mRNA splicing, influencing exon definition and splice-site selection across a broad set of transcripts. Through coupling of RNA processing with transcription and mRNA export, SRp20 helps coordinate gene-expression programs that control cell-cycle progression, differentiation, and stress responses. Its activity contributes to pathway-level regulation in RNA metabolism networks, including spliceosome function and post-transcriptional control of transcripts encoding signaling and chromatin regulators. Dysregulated SRSF3 expression or splicing activity has been associated with aberrant isoform landscapes observed in cancer and other disorders marked by altered RNA processing, supporting its relevance for mechanistic studies of splicing-dependent phenotypes.

SRp20 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SRSF3 expression without altering the underlying DNA sequence.

SRp20 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SRSF3 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 SRSF3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SRp20 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SRSF3 locus and enabling the study of SRp20-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SRp20 pathway restoration in tumor cells with silenced or reduced SRSF3 expression.

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