SRPK1 CRISPR Activation Plasmid (h): sc-402855-ACT

SRPK1 (serine/arginine-rich protein-specific kinase 1) is a human kinase that phosphorylates SR splicing factors to regulate spliceosome assembly, alternative pre-mRNA splicing, and mRNA export. By coupling phosphorylation-dependent localization of splicing regulators with transcriptional outputs, SRPK1 helps shape isoform programs that influence cell-cycle progression, stress responses, and signaling network plasticity. Dysregulated SRPK1 activity and splicing patterns have been linked to altered angiogenic and oncogenic transcript isoforms in cancer biology, and to broader RNA-processing defects observed across neurological and metabolic disease contexts.

SRPK1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SRPK1 expression without altering the underlying DNA sequence.

SRPK1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SRPK1 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 SRPK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SRPK1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SRPK1 locus and enabling the study of SRPK1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SRPK1 pathway restoration in tumor cells with silenced or reduced SRPK1 expression.

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