HSPC171 CRISPR Activation Plasmid (h): sc-412030-ACT

TMEM208 encodes the human membrane protein HSPC171, an endoplasmic reticulum–associated factor implicated in maintaining ER homeostasis and coordinating adaptive responses to proteostatic and metabolic stress. By influencing ER organization and signaling outputs linked to unfolded protein response pathways, TMEM208 can affect downstream processes such as protein quality control, lipid handling, and stress-induced transcriptional programs. Altered ER stress signaling and proteostasis imbalance are recurrent features of cancer, neurodegeneration, and metabolic disorders, making TMEM208/HSPC171 relevant for mechanistic studies that connect ER function to cell survival, differentiation, and inflammatory signaling. As a transmembrane component with potential regulatory roles, TMEM208 is also of interest for mapping organelle-resident networks and identifying context-specific dependencies across cell types.

HSPC171 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM208 expression without altering the underlying DNA sequence.

HSPC171 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM208 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 TMEM208 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSPC171 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM208 locus and enabling the study of HSPC171-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSPC171 pathway restoration in tumor cells with silenced or reduced TMEM208 expression.

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