TMEM38B CRISPR Activation Plasmid (h): sc-412461-ACT

TMEM38B encodes trimeric intracellular cation channel A (TRIC-A), an endoplasmic/sarcoplasmic reticulum membrane protein that forms monovalent cation channels supporting Ca2+ release by providing counter-ion flux. By stabilizing ER/SR membrane potential during IP3 receptor- and ryanodine receptor–dependent calcium signaling, TMEM38B contributes to excitation–contraction coupling, intracellular calcium homeostasis, and downstream calcium-regulated transcriptional programs. Disruption of TMEM38B function has been linked to skeletal development and connective tissue integrity, with loss-of-function variants associated with osteogenesis imperfecta–like phenotypes. These features make TMEM38B relevant for mechanistic studies of ER calcium handling, matrix biology, and stress-associated signaling responses in human cells.

TMEM38B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM38B expression without altering the underlying DNA sequence.

TMEM38B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM38B 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 TMEM38B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM38B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM38B locus and enabling the study of TMEM38B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM38B pathway restoration in tumor cells with silenced or reduced TMEM38B expression.

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