β3Gn-T1 CRISPR Activation Plasmid (h): sc-411554-ACT

B4GAT1 encodes β3Gn-T1, a glycosyltransferase that supports synthesis and remodeling of glycoconjugates by catalyzing β1,3-N-acetylglucosamine transfer reactions that contribute to glycan chain extension. Through its role in protein and lipid glycosylation, β3Gn-T1 influences membrane trafficking, receptor organization, and extracellular matrix interactions that shape cell–cell communication and adhesion-dependent signaling. Altered glycosylation programs involving B4GAT1 have been investigated in contexts of developmental regulation and disease-associated changes in glycan composition, where shifts in surface glycoproteins can impact immune recognition and signaling networks. These properties make B4GAT1 a useful target for studying glycosylation-dependent pathway modulation and phenotypes linked to cell-surface glycans.

β3Gn-T1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous B4GAT1 expression without altering the underlying DNA sequence.

β3Gn-T1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the B4GAT1 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 B4GAT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β3Gn-T1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native B4GAT1 locus and enabling the study of β3Gn-T1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of β3Gn-T1 pathway restoration in tumor cells with silenced or reduced B4GAT1 expression.

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