PACE4 CRISPR Activation Plasmid (h2): sc-405605-ACT-2

Human PCSK6 encodes the proprotein convertase PACE4, a secreted and extracellular matrix–associated serine endoprotease that activates diverse precursor proteins by cleaving paired basic residues within the secretory pathway and at the cell surface. PACE4 contributes to proteostasis and intercellular signaling by processing growth factors, receptors, metalloproteinases, and extracellular matrix components, thereby influencing pathways linked to development, tissue remodeling, and cell migration. Dysregulated PCSK6/PACE4 activity has been associated with altered proteolytic networks in cancer progression, cardiovascular traits, and inflammatory or fibrotic remodeling contexts. PCSK6 gene editing and functional perturbation tools enable mechanistic studies of convertase-dependent substrate maturation, secretome profiling, and dissection of protease-regulated signaling circuits in human cell models.

PACE4 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous PCSK6 expression without altering the underlying DNA sequence.

PACE4 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PCSK6 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 PCSK6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PACE4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PCSK6 locus and enabling the study of PACE4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PACE4 pathway restoration in tumor cells with silenced or reduced PCSK6 expression.

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