GalNAc-T6 CRISPR Activation Plasmid (m): sc-431525-ACT

Galnt6 encodes polypeptide N-acetylgalactosaminyltransferase 6 (GalNAc-T6), a Golgi-resident initiating enzyme for mucin-type O-glycosylation that transfers GalNAc to Ser/Thr residues on secreted and membrane proteins. By establishing O-glycan sites on mucins and other glycoproteins, GalNAc-T6 influences protein folding, protease susceptibility, receptor organization, and extracellular matrix interactions that shape cell adhesion, motility, and signal transduction. Altered O-glycosylation patterns are frequently linked to epithelial remodeling and inflammatory microenvironments, making Galnt6 a relevant node for studying glycoproteome regulation in disease-associated contexts. In mouse models, modulation of Galnt6 activity can be used to interrogate how initiation of O-glycosylation impacts tissue homeostasis and pathway responsiveness.

GalNAc-T6 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Galnt6 expression without altering the underlying DNA sequence.

GalNAc-T6 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Galnt6 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 Galnt6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GalNAc-T6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Galnt6 locus and enabling the study of GalNAc-T6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GalNAc-T6 pathway restoration in tumor cells with silenced or reduced Galnt6 expression.

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