HPRG CRISPR Activation Plasmid (h): sc-404772-ACT

Histidine-rich glycoprotein (HPRG), encoded by the human HRG gene, is a secreted plasma protein that binds heparin, plasminogen, fibrin(ogen), and divalent metal ions to modulate extracellular matrix interactions and proteolytic balance. Through these interactions, HPRG influences coagulation and fibrinolysis dynamics, angiogenic signaling, and innate immune processes including neutrophil behavior and clearance of cellular debris. HRG expression and circulating HPRG levels have been investigated as variables linked to inflammatory states and vascular biology, with implications for interpreting hemostasis- and immunity-associated phenotypes. As a multifunctional ligand-binding protein, HPRG is frequently studied in pathways connecting thromboinflammation, matrix remodeling, and cell–surface glycosaminoglycan signaling.

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

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

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