NDST1 CRISPR Activation Plasmid (h): sc-404078-ACT

NDST1 encodes N-deacetylase/N-sulfotransferase 1, a Golgi-resident enzyme that catalyzes key modification steps during heparan sulfate biosynthesis, including N-deacetylation and N-sulfation of glucosamine residues. These reactions shape heparan sulfate sulfation patterns that determine binding and signaling outputs for growth factors, morphogens, and chemokines across extracellular matrix and cell-surface proteoglycans. Through modulation of heparan sulfate–dependent pathways such as FGF, Wnt, Hedgehog, and BMP signaling, NDST1 influences cell adhesion, migration, differentiation, and tissue patterning. Dysregulated heparan sulfate remodeling has been linked to altered developmental programs and microenvironmental signaling relevant to cancer biology, inflammation, and neurological and vascular phenotypes, motivating mechanistic studies of NDST1-controlled glycosaminoglycan architecture.

NDST1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NDST1 expression without altering the underlying DNA sequence.

NDST1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NDST1 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 NDST1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NDST1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NDST1 locus and enabling the study of NDST1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NDST1 pathway restoration in tumor cells with silenced or reduced NDST1 expression.

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