GlcAT-S CRISPR Activation Plasmid (h): sc-408633-ACT

B3GAT2 encodes glucuronyltransferase S (GlcAT-S), a Golgi-localized glycosyltransferase that transfers glucuronic acid during biosynthesis of the glycosaminoglycan-protein linkage region required for proteoglycan assembly. Through its role in glycosylation and extracellular matrix organization, GlcAT-S influences cell–matrix interactions, receptor signaling, and developmental processes dependent on properly modified proteoglycans. Perturbation of proteoglycan biosynthesis and linkage-region formation is linked to altered tissue architecture and signaling abnormalities relevant to connective tissue and neurobiology research. As a human glycoenzyme, GlcAT-S is also of interest for studying how Golgi glycosylation states shape cell surface composition and downstream pathway activity.

GlcAT-S CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous B3GAT2 expression without altering the underlying DNA sequence.

GlcAT-S CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the B3GAT2 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 B3GAT2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GlcAT-S expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native B3GAT2 locus and enabling the study of GlcAT-S-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GlcAT-S pathway restoration in tumor cells with silenced or reduced B3GAT2 expression.

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