TMEM214 CRISPR Activation Plasmid (m2): sc-427183-ACT-2

Mouse Tmem214 encodes TMEM214, an endoplasmic reticulum–associated transmembrane protein implicated in the regulation of apoptosis and ER stress signaling. TMEM214 has been linked to modulation of the unfolded protein response and Ca2+ homeostasis, influencing cell survival decisions under proteotoxic stress and perturbations in ER function. Dysregulation of ER stress and apoptotic pathways is relevant to mechanisms studied in neurodegeneration, metabolic dysfunction, and cancer biology, making Tmem214 a useful target for pathway dissection. Gene editing of Tmem214 in murine cells or in vivo models enables functional interrogation of ER stress–apoptosis crosstalk, downstream transcriptional programs, and stress-adaptive phenotypes in biomedical research.

TMEM214 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Tmem214 expression without altering the underlying DNA sequence.

TMEM214 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Tmem214 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 Tmem214 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM214 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Tmem214 locus and enabling the study of TMEM214-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM214 pathway restoration in tumor cells with silenced or reduced Tmem214 expression.

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