OX2R CRISPR Activation Plasmid (h): sc-404121-ACT

Human CD200R1 encodes the inhibitory immunoreceptor OX2R, a myeloid-lineage checkpoint receptor that binds CD200 to restrain inflammatory activation. OX2R signaling dampens cytokine production and antigen-presenting cell maturation through phosphorylation-dependent inhibitory pathways that converge on reduced MAPK and NF-κB activity, shaping innate–adaptive immune crosstalk. This axis is implicated in regulation of microglial homeostasis and peripheral macrophage responses, with relevance to immune evasion and chronic inflammatory states observed in cancer, autoimmune conditions, and neuroinflammatory disease models. As a result, CD200R1 is frequently studied in pathways controlling immune tolerance, tissue damage responses, and tumor-associated myeloid phenotypes.

OX2R CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD200R1 expression without altering the underlying DNA sequence.

OX2R CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD200R1 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 CD200R1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OX2R expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD200R1 locus and enabling the study of OX2R-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OX2R pathway restoration in tumor cells with silenced or reduced CD200R1 expression.

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