AGR3 CRISPR Activation Plasmid (h): sc-406892-ACT

Anterior gradient protein 3 (AGR3) is a human protein disulfide isomerase family member localized to the secretory pathway, where it supports oxidative protein folding and maturation of select client proteins. AGR3 contributes to endoplasmic reticulum proteostasis and epithelial differentiation programs, and its expression is often enriched in mucus-producing and secretory epithelia. Dysregulated AGR3 expression has been reported in multiple epithelial malignancies and is studied in the context of tumor cell phenotype, stress adaptation, and secretory pathway remodeling. These features make AGR3 a useful node for investigating ER quality control, secretion-linked signaling, and transcriptional states associated with epithelial biology.

AGR3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AGR3 expression without altering the underlying DNA sequence.

AGR3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AGR3 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 AGR3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AGR3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AGR3 locus and enabling the study of AGR3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AGR3 pathway restoration in tumor cells with silenced or reduced AGR3 expression.

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