TMEM97 CRISPR Activation Plasmid (h): sc-411997-ACT

TMEM97 encodes a small multi-pass transmembrane protein also known as the sigma-2 receptor, enriched in endoplasmic reticulum and lysosomal/endosomal membranes and implicated in intracellular cholesterol handling and membrane trafficking. TMEM97 has been linked to regulation of sterol distribution through interactions with lipid transport and receptor complexes, influencing vesicular dynamics and cellular stress responses. Altered TMEM97 expression has been reported across proliferative and neurodegenerative contexts, where changes in lipid homeostasis, oxidative stress, and proteostasis can contribute to disease-associated phenotypes. These attributes make TMEM97 a useful target for mechanistic studies of lipid-regulated signaling, organelle function, and cell-state transitions.

TMEM97 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM97 expression without altering the underlying DNA sequence.

TMEM97 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM97 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 TMEM97 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM97 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM97 locus and enabling the study of TMEM97-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM97 pathway restoration in tumor cells with silenced or reduced TMEM97 expression.

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