TMEM132A CRISPR Activation Plasmid (h): sc-412402-ACT

TMEM132A (transmembrane protein 132A) is a human single-pass membrane protein implicated in cell–cell communication and regulation of adhesion-related signaling at the cell surface. Expression of TMEM132A has been linked to neural and glial biology, including pathways that influence cytoskeletal organization and membrane-associated signaling processes that shape cell morphology and connectivity. Genetic and expression studies have associated TMEM132A with neuropsychiatric and neurodevelopmental phenotypes, supporting its relevance for investigating mechanisms of neuronal function and circuit organization. As a membrane-associated factor, TMEM132A is also useful for probing how extracellular cues are translated into intracellular responses that impact differentiation and stress responsiveness.

TMEM132A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM132A expression without altering the underlying DNA sequence.

TMEM132A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM132A 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 TMEM132A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM132A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM132A locus and enabling the study of TMEM132A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM132A pathway restoration in tumor cells with silenced or reduced TMEM132A expression.

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