CRP CRISPR Activation Plasmid (h): sc-401767-ACT

C-reactive protein (CRP) is a secreted pentraxin produced primarily by hepatocytes as part of the acute-phase response, and it functions as a soluble pattern-recognition molecule that binds phosphocholine on microbial surfaces and damaged cells. By engaging Fcγ receptors and activating the classical complement cascade via C1q, CRP links innate immune sensing to opsonization, phagocytosis, and inflammatory amplification. CRP expression is transcriptionally induced by pro-inflammatory cytokines, especially IL-6 with contributions from IL-1β and TNF, integrating JAK/STAT and NF-κB-driven signaling programs. Dysregulated CRP levels are widely used as a biomarker of systemic inflammation and are associated with cardiometabolic disease, infection, and autoimmune pathophysiology, supporting mechanistic studies of inflammatory regulation.

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

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

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