Fibrinogen β CRISPR Activation Plasmid (h): sc-401819-ACT

FGB encodes the fibrinogen β chain, an essential component of the fibrinogen hexamer that is proteolytically converted by thrombin into fibrin during the terminal phase of the coagulation cascade. Along with interactions with platelet integrin αIIbβ3 and other extracellular matrix proteins, fibrinogen supports clot formation, wound repair, and modulation of inflammatory cell recruitment. FGB expression is tightly linked to hepatic acute-phase responses and cytokine signaling programs that reshape plasma protein composition during systemic inflammation. Genetic or acquired perturbations in fibrinogen structure or abundance have been associated with bleeding or thrombotic phenotypes, making FGB a useful node for studying hemostasis and inflammation-associated vascular pathology.

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

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

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