TMEM167B CRISPR Activation Plasmid (h): sc-410050-ACT

TMEM167B encodes a multi-pass transmembrane protein implicated in membrane trafficking and endomembrane system organization, with reported enrichment in secretory and vesicular compartments. Emerging studies link TMEM167B to regulation of cellular homeostasis processes such as protein sorting and organelle-associated signaling, which can influence receptor availability and downstream pathway responsiveness. Altered expression patterns have been observed across multiple omics datasets, supporting continued investigation of TMEM167B in contexts including cell stress adaptation and dysregulated growth programs. As a relatively understudied membrane protein, TMEM167B is well suited for mechanistic research aimed at mapping trafficking-dependent phenotypes and identifying genetic interactions.

TMEM167B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM167B expression without altering the underlying DNA sequence.

TMEM167B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM167B 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 TMEM167B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM167B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM167B locus and enabling the study of TMEM167B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM167B pathway restoration in tumor cells with silenced or reduced TMEM167B expression.

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