Selenoprotein I CRISPR Activation Plasmid (h): sc-408993-ACT

Human SELENOI encodes selenoprotein I, an endoplasmic reticulum–localized enzyme that functions as ethanolamine phosphotransferase, catalyzing a key step in de novo phosphatidylethanolamine biosynthesis. By controlling membrane phospholipid composition, SELENOI influences ER homeostasis, vesicular trafficking, and lipid-dependent signaling processes that shape cellular stress responses and organelle function. Perturbations in ethanolamine phospholipid metabolism have been linked to altered membrane dynamics and mitochondrial-ER communication, processes relevant to neurobiology and metabolic regulation. As a selenoprotein, SELENOI also intersects with redox-sensitive pathways through selenium biology and ER-associated protein quality control.

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

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

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