SFRS2B CRISPR Activation Plasmid (h): sc-401973-ACT

SRSF8 encodes the serine/arginine-rich splicing factor SFRS2B, an RNA-binding protein that participates in spliceosome assembly and regulates alternative pre-mRNA splicing decisions. Through recognition of exonic splicing enhancers and coordination of exon inclusion or skipping, SFRS2B influences mRNA isoform diversity, transcript stability, and downstream proteome composition. Splicing-factor activity intersects with transcriptional elongation, mRNA processing, and stress-responsive RNA metabolism, shaping cell-state programs such as proliferation and differentiation. Dysregulated alternative splicing and altered expression of SR-family regulators are recurrent features of human disease biology, supporting investigation of SRSF8-dependent isoform networks in oncogenic and neurobiological contexts.

SFRS2B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SRSF8 expression without altering the underlying DNA sequence.

SFRS2B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SRSF8 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 SRSF8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SFRS2B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SRSF8 locus and enabling the study of SFRS2B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SFRS2B pathway restoration in tumor cells with silenced or reduced SRSF8 expression.

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