OX2R CRISPR Activation Plasmid (m): sc-425439-ACT

Mouse Cd200r1 encodes OX2R (CD200 receptor 1), an inhibitory immunoregulatory receptor expressed predominantly by myeloid lineage cells including macrophages, dendritic cells, and microglia. Engagement of OX2R by its ligand CD200 promotes ITIM-dependent signaling that recruits phosphatases such as SHP-1/2 to dampen pro-inflammatory cascades, thereby constraining cytokine production, phagocyte activation, and antigen-presenting functions. Through these pathways, Cd200r1 contributes to maintenance of immune tolerance and tissue homeostasis at barrier sites and within the central nervous system. Dysregulation of CD200–OX2R signaling has been implicated in models of neuroinflammation, autoimmunity, infection-associated immunopathology, and tumor immune evasion, making it a useful node for dissecting myeloid checkpoint biology.

OX2R CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cd200r1 expression without altering the underlying DNA sequence.

OX2R CRISPR Activation Plasmid (m) 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.