Fibronectin CRISPR Activation Plasmid (h): sc-400082-ACT

FN1 encodes fibronectin, a major extracellular matrix glycoprotein that orchestrates cell adhesion, migration, and tissue remodeling through binding to integrins, collagen, and heparan sulfate proteoglycans. Fibronectin participates in focal adhesion and mechanotransduction signaling, coordinating actin cytoskeleton dynamics and regulating pathways such as FAK/SRC, Rho GTPase signaling, and TGF-β–driven extracellular matrix organization. In human biology, altered FN1 expression or deposition is associated with dysregulated wound repair, fibrosis, tumor cell invasion, and remodeling of the stromal microenvironment. These properties make FN1 a central target for studying extracellular matrix assembly, epithelial–mesenchymal plasticity, and integrin-dependent signaling networks.

Fibronectin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FN1 expression without altering the underlying DNA sequence.

Fibronectin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FN1 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 FN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fibronectin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FN1 locus and enabling the study of Fibronectin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fibronectin pathway restoration in tumor cells with silenced or reduced FN1 expression.

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