TF CRISPR Activation Plasmid (h): sc-401060-ACT

Human F3 encodes tissue factor (TF), a transmembrane glycoprotein that initiates the extrinsic coagulation cascade by binding factor VII/VIIa and promoting downstream thrombin generation and fibrin formation. Beyond hemostasis, TF influences cell signaling through protease-activated receptor pathways and can modulate inflammation, endothelial activation, and vascular integrity. F3 expression is regulated by cytokine- and stress-responsive programs, linking TF activity to immune–coagulation crosstalk and tissue injury responses. Dysregulated TF levels and signaling have been associated with thromboinflammatory states and tumor-associated procoagulant activity, making F3 a useful target for mechanistic studies in vascular biology and cancer-associated pathways.

TF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous F3 expression without altering the underlying DNA sequence.

TF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the F3 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 F3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native F3 locus and enabling the study of TF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TF pathway restoration in tumor cells with silenced or reduced F3 expression.

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