TMEM215 CRISPR Activation Plasmid (h2): sc-408002-ACT-2

Human TMEM215 encodes transmembrane protein 215, a predicted multi-pass membrane protein thought to localize to cellular membranes where it may contribute to membrane organization, transport-associated processes, or signaling-dependent regulation of cellular homeostasis. Although TMEM215 remains incompletely characterized, expression and genetic association data have linked it to pathways relevant to cell proliferation, differentiation, and stress responses, suggesting context-dependent roles in tissue physiology. Altered TMEM215 expression patterns have been reported in disease-related datasets, including cancer and other complex disorders, supporting its utility as a target for functional genomics studies of membrane protein biology. Gene editing of TMEM215 enables loss- or gain-of-function interrogation of subcellular localization, protein–protein interactions, and downstream transcriptional or phenotypic effects in human cell models.

TMEM215 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous TMEM215 expression without altering the underlying DNA sequence.

TMEM215 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM215 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 TMEM215 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM215 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM215 locus and enabling the study of TMEM215-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM215 pathway restoration in tumor cells with silenced or reduced TMEM215 expression.

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