IFN-α/βRα CRISPR Activation Plasmid (h): sc-401662-ACT

IFNAR1 encodes interferon-α/β receptor alpha (IFN-α/βRα), an essential cell-surface subunit of the type I interferon receptor complex that binds IFN-α and IFN-β to initiate antiviral and immunomodulatory signaling. Ligand engagement activates JAK1/TYK2 and downstream STAT1/STAT2–IRF9 (ISGF3) transcriptional programs, inducing interferon-stimulated genes that shape innate immunity, antigen presentation, and inflammatory tone. IFNAR1-dependent pathways intersect with NF-κB and MAPK signaling and influence cell survival, differentiation, and cytokine networks across immune and stromal compartments. Dysregulated IFNAR1 signaling is implicated in altered antiviral responses and interferon-driven immune phenotypes relevant to infection biology, autoimmunity-associated signatures, and tumor–immune interactions.

IFN-α/βRα CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFNAR1 expression without altering the underlying DNA sequence.

IFN-α/βRα CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFNAR1 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 IFNAR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFN-α/βRα expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFNAR1 locus and enabling the study of IFN-α/βRα-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFN-α/βRα pathway restoration in tumor cells with silenced or reduced IFNAR1 expression.

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