CHST1 CRISPR Activation Plasmid (h): sc-403978-ACT

CHST1 encodes carbohydrate sulfotransferase 1, a Golgi-resident enzyme that transfers sulfate to galactose residues on glycoproteins and glycolipids, contributing to the biosynthesis of sulfated glycosaminoglycans and related glycoconjugates. By shaping cell-surface and extracellular matrix sulfation patterns, CHST1 influences ligand–receptor interactions, leukocyte adhesion, and growth factor/cytokine signaling within glycosylation and proteoglycan metabolism pathways. Altered sulfation can remodel the glycocalyx and modulate immune recognition and cell migration, linking CHST1 activity to mechanisms relevant to inflammation and tumor cell behavior. CHST1 is therefore of interest for studies of glycan-dependent regulation of signaling, adhesion, and microenvironmental interactions.

CHST1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHST1 expression without altering the underlying DNA sequence.

CHST1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHST1 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 CHST1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CHST1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHST1 locus and enabling the study of CHST1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CHST1 pathway restoration in tumor cells with silenced or reduced CHST1 expression.

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