MRGX2 CRISPR Activation Plasmid (h): sc-414107-ACT

MRGPRX2 encodes the human Mas-related G protein-coupled receptor X2 (MRGX2), a GPCR highly enriched in mast cells that mediates IgE-independent activation in response to diverse cationic peptides and small molecules. Receptor engagement triggers Gαq/PLC signaling, intracellular Ca2+ mobilization, and downstream MAPK and NF-κB-linked transcriptional programs that shape degranulation and proinflammatory mediator release. MRGX2 has been implicated in neuroimmune communication and pseudo-allergic reactions, and its dysregulation is studied in the context of chronic urticaria, atopic inflammation, and drug-induced immediate hypersensitivity-like responses. These properties make MRGPRX2 a useful node for interrogating mast cell activation thresholds, receptor-biased signaling, and inflammatory pathway crosstalk.

MRGX2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MRGPRX2 expression without altering the underlying DNA sequence.

MRGX2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MRGPRX2 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 MRGPRX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRGX2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MRGPRX2 locus and enabling the study of MRGX2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRGX2 pathway restoration in tumor cells with silenced or reduced MRGPRX2 expression.

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