Stomatin CRISPR Activation Plasmid (h): sc-402491-ACT

Human STOM encodes stomatin, an integral membrane protein enriched in lipid rafts that modulates membrane organization and ion channel activity, including regulation of mechanosensitive and epithelial sodium channel complexes. Stomatin contributes to cytoskeletal coupling, vesicular trafficking, and membrane microdomain-dependent signaling that influence cell volume control and stress responses. Altered STOM expression or membrane localization has been associated with red blood cell membrane defects such as hereditary stomatocytosis and can impact cellular biomechanics and ion homeostasis. As a raft-associated scaffold, stomatin is frequently studied in pathways linking membrane tension sensing, ion transport, and inflammatory or metabolic stress adaptation.

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

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

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