β-1,3-Gal-T5 CRISPR Activation Plasmid (h): sc-409777-ACT

B3GALT5 encodes β-1,3-Gal-T5, a Golgi-resident β1,3-galactosyltransferase that contributes to the synthesis and extension of glycolipids and glycoproteins by adding galactose in β1,3 linkage to N-acetylglucosamine-containing acceptors. Through its role in glycosylation, β-1,3-Gal-T5 influences cell-surface glycan composition that governs protein trafficking, receptor organization, and lectin-mediated cell–cell interactions. Altered B3GALT5 activity can shift glycan-dependent signaling and adhesion programs implicated in immune recognition, epithelial differentiation, and tumor-associated glycosylation phenotypes. As a result, B3GALT5 is a useful target for dissecting how glycosyltransferase networks regulate membrane biology and disease-relevant changes in cell behavior.

β-1,3-Gal-T5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous B3GALT5 expression without altering the underlying DNA sequence.

β-1,3-Gal-T5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the B3GALT5 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 B3GALT5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β-1,3-Gal-T5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native B3GALT5 locus and enabling the study of β-1,3-Gal-T5-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-T5 pathway restoration in tumor cells with silenced or reduced B3GALT5 expression.

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