Selenoprotein S CRISPR Activation Plasmid (h): sc-404444-ACT

SELENOS encodes Selenoprotein S, an endoplasmic reticulum membrane selenoprotein that supports ER-associated degradation (ERAD) and proteostasis by promoting clearance of misfolded proteins. Through modulation of ER stress signaling and redox homeostasis, Selenoprotein S influences unfolded protein response pathways and inflammatory signaling outputs. Variation in SELENOS expression has been linked to altered cytokine production and susceptibility to metabolic and immune-related phenotypes, reflecting its role at the interface of oxidative stress and inflammation. As a human gene involved in ER quality control, SELENOS is frequently studied in contexts of chronic stress adaptation, macrophage activation, and cellular responses to proteotoxic insults.

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

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

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