IFN-ω CRISPR Activation Plasmid (h): sc-409162-ACT

IFNW1 encodes interferon omega (IFN-ω), a type I interferon that contributes to innate antiviral defense and immune surveillance. IFN-ω signals through the IFNAR receptor complex to activate JAK–STAT pathways and induce interferon-stimulated genes (ISGs) that regulate viral restriction, antigen presentation, and modulation of inflammatory programs. Its activity intersects with nucleic acid–sensing pathways such as RIG-I/MDA5 and cGAS–STING that coordinate transcriptional responses to infection and cellular stress. Dysregulated type I interferon signaling, including aberrant IFN-ω expression, is relevant to studies of chronic inflammation, autoimmunity, and tumor–immune interactions where interferon-driven gene networks shape cellular phenotypes.

IFN-omega CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFNW1 expression without altering the underlying DNA sequence.

IFN-omega CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFNW1 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 IFNW1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFN-omega expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFNW1 locus and enabling the study of IFN-omega-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFN-omega pathway restoration in tumor cells with silenced or reduced IFNW1 expression.

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