GCNT3 CRISPR Activation Plasmid (h): sc-411258-ACT

Human GCNT3 (glucosaminyl (N-acetyl) transferase 3) encodes a Golgi-localized glycosyltransferase that extends O-glycans by catalyzing formation of core 2 and core 4 structures on mucin-type glycoproteins. By remodeling glycan branching, GCNT3 influences epithelial differentiation, mucosal barrier properties, and cell–cell or cell–matrix interactions that depend on glycoprotein and glycolipid recognition. Its activity intersects with O-glycosylation networks that shape receptor stability, trafficking, and immune recognition, linking GCNT3 to pathways governing adhesion and inflammatory signaling. Altered GCNT3 expression or glycosylation patterns have been reported in multiple epithelial disease contexts, including cancers, where changes in mucin glycoforms can affect invasion, metastasis-associated phenotypes, and tumor–microenvironment interactions.

GCNT3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GCNT3 expression without altering the underlying DNA sequence.

GCNT3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GCNT3 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 GCNT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GCNT3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GCNT3 locus and enabling the study of GCNT3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GCNT3 pathway restoration in tumor cells with silenced or reduced GCNT3 expression.

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