Biglycan CRISPR Activation Plasmid (h): sc-401408-ACT

BGN encodes biglycan, a secreted small leucine-rich proteoglycan that binds collagens and regulates extracellular matrix (ECM) assembly, tissue mechanics, and pericellular signaling in connective tissues. Biglycan modulates growth factor availability and receptor signaling, intersecting with pathways such as TGF-β/SMAD, Wnt/β-catenin, and integrin-mediated adhesion to influence cell proliferation, migration, and inflammatory responses. In human biology, altered BGN expression and ECM remodeling are frequently associated with fibrotic processes, tumor–stroma interactions, and vascular and musculoskeletal pathophysiology, making it a useful node for studying matrix-driven signaling. Its roles in innate immune activation and cytokine regulation also support mechanistic investigations into inflammation-linked tissue remodeling.

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

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

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