ChemR23 CRISPR Activation Plasmid (h): sc-403033-ACT

CMKLR1 encodes ChemR23, a G protein–coupled receptor that binds chemerin (RARRES2) and the pro-resolving lipid mediator resolvin E1 to regulate leukocyte chemotaxis, adhesion, and innate immune activation. ChemR23 signaling engages Gi-dependent pathways that modulate cAMP, intracellular calcium flux, and downstream MAPK/ERK and PI3K programs, shaping inflammatory tone and tissue remodeling. The CMKLR1 axis is implicated in macrophage and dendritic cell trafficking, adipose tissue biology, and vascular responses, linking receptor activity to chronic inflammatory states. Altered ChemR23 expression and ligand availability have been associated with metabolic inflammation, autoimmune conditions, and tumor microenvironment dynamics, making it a relevant node for pathway interrogation in human cell systems.

ChemR23 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CMKLR1 expression without altering the underlying DNA sequence.

ChemR23 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CMKLR1 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 CMKLR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ChemR23 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CMKLR1 locus and enabling the study of ChemR23-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ChemR23 pathway restoration in tumor cells with silenced or reduced CMKLR1 expression.

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