ORMDL3 CRISPR Activation Plasmid (h): sc-403587-ACT

ORMDL3 (orosomucoid-like 3) is an endoplasmic reticulum membrane protein that functions as a key regulator of sphingolipid homeostasis by modulating serine palmitoyltransferase activity, thereby influencing ceramide production and membrane lipid composition. Through its role in ER lipid metabolism, ORMDL3 impacts calcium signaling, ER stress responses, and inflammatory pathway tone, linking cellular homeostasis to immune activation. Genetic variation and dysregulated expression of ORMDL3 have been associated with asthma susceptibility and other inflammatory phenotypes, supporting its relevance to airway biology and immunometabolism. Researchers commonly interrogate ORMDL3 to understand how lipid-driven signaling and ER proteostasis shape cytokine responses, epithelial function, and stress-adaptive programs.

ORMDL3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ORMDL3 expression without altering the underlying DNA sequence.

ORMDL3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ORMDL3 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 ORMDL3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ORMDL3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ORMDL3 locus and enabling the study of ORMDL3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ORMDL3 pathway restoration in tumor cells with silenced or reduced ORMDL3 expression.

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