EDG-2 CRISPR Activation Plasmid (h): sc-402843-ACT

Human LPAR1 encodes EDG-2, a lysophosphatidic acid (LPA)–responsive G protein-coupled receptor that couples primarily to Gαi, Gαq, and Gα12/13 to regulate MAPK/ERK, PI3K–AKT, PLC–Ca2+ signaling, and Rho/ROCK-dependent cytoskeletal remodeling. Through these pathways, EDG-2 influences cell migration, adhesion, proliferation, survival, and extracellular matrix interactions, integrating lipid mediator cues into tissue remodeling and inflammatory responses. LPAR1 activity is frequently studied in contexts where LPA signaling shapes stromal–immune communication, vascular and epithelial barrier function, and fibroblast activation. Dysregulated EDG-2 signaling has been linked in the literature to cancer-associated invasion, fibrotic remodeling, and neuroinflammatory processes, making it a relevant node for mechanistic pathway dissection.

EDG-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LPAR1 expression without altering the underlying DNA sequence.

EDG-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LPAR1 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 LPAR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EDG-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LPAR1 locus and enabling the study of EDG-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EDG-2 pathway restoration in tumor cells with silenced or reduced LPAR1 expression.

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