β-1,4-GalNAc-T3 CRISPR Activation Plasmid (h): sc-415306-ACT

B4GALNT3 encodes human β-1,4-GalNAc-T3, a Golgi-localized glycosyltransferase that transfers N-acetylgalactosamine in β1,4 linkage to generate specific terminal glycan epitopes on glycoproteins and glycolipids. By shaping cell-surface and secreted glycosylation patterns, β-1,4-GalNAc-T3 influences protein trafficking, receptor–ligand interactions, and lectin-mediated signaling that impact cell adhesion and communication. Altered B4GALNT3 expression or activity has been associated with dysregulated glycan remodeling observed in cancer biology and other conditions where aberrant glycosylation modulates proliferation, invasion, and immune recognition. This gene is therefore relevant for studies of glycosylation-dependent pathways, glycocalyx composition, and mechanisms linking Golgi enzyme programs to disease phenotypes.

β-1,4-GalNAc-T3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous B4GALNT3 expression without altering the underlying DNA sequence.

β-1,4-GalNAc-T3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the B4GALNT3 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 B4GALNT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β-1,4-GalNAc-T3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native B4GALNT3 locus and enabling the study of β-1,4-GalNAc-T3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of β-1,4-GalNAc-T3 pathway restoration in tumor cells with silenced or reduced B4GALNT3 expression.

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