EDEM3 CRISPR Activation Plasmid (h): sc-408476-ACT

EDEM3 (ER degradation enhancing alpha-mannosidase-like protein 3) is an ER-resident component of glycoprotein quality control that participates in ER-associated degradation (ERAD) by promoting demannosylation-dependent recognition and clearance of misfolded N-glycoproteins. By influencing the unfolded protein response, proteostasis, and secretory pathway fidelity, EDEM3 helps regulate cellular stress adaptation and turnover of aberrant client proteins. Perturbation of ERAD and ER proteostasis is broadly relevant to disease biology, including contexts where chronic ER stress, altered glycan processing, and dysregulated protein degradation contribute to pathogenesis. Human EDEM3 is therefore a useful node for studying ER quality control circuitry and secretory pathway homeostasis in mechanistic and functional genomics workflows.

EDEM3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EDEM3 expression without altering the underlying DNA sequence.

EDEM3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EDEM3 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 EDEM3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EDEM3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EDEM3 locus and enabling the study of EDEM3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EDEM3 pathway restoration in tumor cells with silenced or reduced EDEM3 expression.

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