ERp57 CRISPR Activation Plasmid (h): sc-401497-ACT

PDIA3 encodes ERp57, an endoplasmic reticulum (ER) protein disulfide isomerase that cooperates with calnexin and calreticulin to catalyze disulfide bond formation and isomerization during glycoprotein folding. ERp57 contributes to ER proteostasis, quality control, and ER-associated processes linked to the unfolded protein response and oxidative protein folding. Beyond the ER lumen, PDIA3 has been implicated in antigen presentation through MHC class I peptide loading and broader regulation of cellular stress signaling. Dysregulated ERp57 activity or expression is associated with altered protein homeostasis and has been studied in the context of inflammation, neurodegeneration, and cancer-related pathways.

ERp57 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PDIA3 expression without altering the underlying DNA sequence.

ERp57 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PDIA3 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 PDIA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ERp57 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PDIA3 locus and enabling the study of ERp57-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ERp57 pathway restoration in tumor cells with silenced or reduced PDIA3 expression.

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