β-1,3-Gal-T3 CRISPR Activation Plasmid (h): sc-411200-ACT

B3GALNT1 encodes human β-1,3-Gal-T3, a Golgi-localized glycosyltransferase that catalyzes transfer of N-acetylgalactosamine to galactose residues to generate specific glycan structures on glycolipids and glycoproteins. Through its role in glycosylation, β-1,3-Gal-T3 influences membrane organization, receptor behavior, and cell–cell communication by shaping glycan-dependent recognition and trafficking. Altered expression or activity of B3GALNT1 can perturb glycosphingolipid and glycoprotein biosynthesis, with downstream effects on adhesion, migration, and signaling networks. Dysregulated glycosylation patterns involving B3GALNT1 have been investigated in contexts where glycan remodeling is linked to disease-associated phenotypes and biomarker changes.

β-1,3-Gal-T3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous B3GALNT1 expression without altering the underlying DNA sequence.

β-1,3-Gal-T3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the B3GALNT1 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 B3GALNT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β-1,3-Gal-T3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native B3GALNT1 locus and enabling the study of β-1,3-Gal-T3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of β-1,3-Gal-T3 pathway restoration in tumor cells with silenced or reduced B3GALNT1 expression.

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