SAP CRISPR Activation Plasmid (h): sc-402165-ACT

APCS encodes serum amyloid P component (SAP), a pentraxin family glycoprotein that binds calcium-dependently to microbial polysaccharides, nuclear debris, and amyloid fibrils, influencing opsonization and complement-related innate immune processes. SAP participates in pattern recognition and clearance pathways by interacting with Fc receptors and complement components, shaping inflammatory signaling and tissue homeostasis. Altered SAP abundance or binding activity has been associated with amyloid deposition biology and dysregulated inflammatory states, making APCS a useful target for studying extracellular matrix-associated protein complexes and immune surveillance mechanisms. In human systems, SAP is also used as a molecular readout for pentraxin-mediated modulation of soluble ligand binding and aggregation-prone protein handling.

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

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

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