RISC CRISPR Activation Plasmid (h): sc-406937-ACT

Human SCPEP1 (serine carboxypeptidase 1) encodes a secreted/extracellular serine carboxypeptidase–like protein implicated in proteolytic processing and remodeling of the pericellular microenvironment. SCPEP1 has been linked to cellular programs involved in vascular biology, including smooth muscle cell phenotype modulation, migration, and responses to inflammatory cues that intersect with extracellular matrix turnover and growth factor signaling. Altered SCPEP1 expression has been reported in contexts of vascular remodeling and cardiometabolic disease–relevant processes, supporting its use as a mechanistic node in studies of atherosclerosis-associated pathways. As part of secreted protease networks, SCPEP1 is also of interest for dissecting how extracellular proteolysis influences cell–cell communication and tissue homeostasis.

RISC CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SCPEP1 expression without altering the underlying DNA sequence.

RISC CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SCPEP1 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 SCPEP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RISC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SCPEP1 locus and enabling the study of RISC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RISC pathway restoration in tumor cells with silenced or reduced SCPEP1 expression.

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