ZAG CRISPR Activation Plasmid (h): sc-401706-ACT

AZGP1 encodes zinc-α2-glycoprotein (ZAG), a secreted glycoprotein that shares structural homology with MHC class I proteins and is detected in multiple epithelial tissues and body fluids. ZAG is implicated in lipid mobilization and metabolic regulation, influencing adipocyte biology and systemic energy balance through pathways linked to fatty-acid utilization and cellular stress responses. Altered AZGP1/ZAG expression has been reported across diverse contexts including epithelial tumor biology, cachexia-associated metabolic remodeling, and inflammatory states, supporting its use as a molecular readout of tissue differentiation and microenvironmental signaling. Because ZAG is extracellular and measurable, it is also commonly studied as a biomarker-like protein in mechanistic studies of secretion, glycosylation, and intercellular communication.

ZAG CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AZGP1 expression without altering the underlying DNA sequence.

ZAG CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AZGP1 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 AZGP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZAG expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AZGP1 locus and enabling the study of ZAG-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZAG pathway restoration in tumor cells with silenced or reduced AZGP1 expression.

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