IFN-α/βRβ CRISPR Activation Plasmid (h): sc-403854-ACT

IFNAR2 encodes the interferon alpha/beta receptor subunit 2 (IFN-α/βRβ), a core component of the type I interferon receptor complex that mediates cellular responses to IFN-α and IFN-β. Ligand engagement activates JAK1/TYK2-dependent signaling and downstream STAT1/STAT2-IRF9 (ISGF3) transcriptional programs, coordinating antiviral defense, antigen presentation, and innate immune regulation. IFNAR2-dependent pathways modulate cross-talk with NF-κB and MAPK signaling and influence cytokine networks that shape immune cell activation and tissue inflammation. Altered IFNAR2 expression or signaling capacity is frequently investigated in contexts of dysregulated interferon-stimulated gene expression, immune-mediated pathology, and tumor immune interactions.

IFN-α/βRβ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFNAR2 expression without altering the underlying DNA sequence.

IFN-α/βRβ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFNAR2 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 IFNAR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFN-α/βRβ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFNAR2 locus and enabling the study of IFN-α/βRβ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFN-α/βRβ pathway restoration in tumor cells with silenced or reduced IFNAR2 expression.

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