brevican CRISPR Activation Plasmid (h): sc-402668-ACT

BCAN encodes brevican, a chondroitin sulfate proteoglycan enriched in the brain extracellular matrix and perineuronal nets, where it modulates cell–matrix interactions and the structural plasticity of synapses. Brevican binds hyaluronan and associates with tenascin-R and other matrix constituents to influence extracellular matrix assembly, neuronal adhesion, and neurite outgrowth. Through regulation of pericellular matrix organization and protease-sensitive remodeling, BCAN contributes to processes linked to neurodevelopment and activity-dependent circuit refinement. Altered brevican expression and processing has been reported in neurological and neuropsychiatric contexts and is frequently evaluated as a matrix-associated factor in glioma biology and invasive phenotypes.

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

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

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