ORMDL2 CRISPR Activation Plasmid (h): sc-403585-ACT

ORMDL2 (orosomucoid-like 2) is an endoplasmic reticulum membrane protein that participates in sphingolipid homeostasis by modulating serine palmitoyltransferase activity and balancing ceramide and sphingomyelin synthesis. Through this role, ORMDL2 influences ER membrane composition, calcium handling, and ER stress signaling, linking lipid metabolism to proteostasis and inflammatory responses. Altered ORMDL family regulation has been associated with airway inflammation, asthma-related traits, and broader immunometabolic phenotypes, making ORMDL2 a useful target for studying lipid-driven signaling in human cells. Its position at the intersection of sphingolipid biosynthesis and stress-response pathways supports mechanistic research into how ER lipid flux shapes innate immune and barrier functions.

ORMDL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ORMDL2 expression without altering the underlying DNA sequence.

ORMDL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ORMDL2 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 ORMDL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ORMDL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ORMDL2 locus and enabling the study of ORMDL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ORMDL2 pathway restoration in tumor cells with silenced or reduced ORMDL2 expression.

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