IFN-β CRISPR Activation Plasmid (h): sc-418564-ACT

IFNB1 encodes human interferon‑β (IFN‑β), a type I interferon that is rapidly induced downstream of pattern recognition receptors sensing viral and cytosolic nucleic acids. Secreted IFN‑β activates IFNAR-dependent JAK–STAT signaling to drive transcription of interferon-stimulated genes, shaping antiviral defense, antigen presentation, and innate-adaptive immune crosstalk. IFNB1 is integrated with IRF3/IRF7, NF‑κB, and cGAS–STING/TBK1 signaling nodes that coordinate inflammatory gene programs and cellular stress responses. Dysregulated IFN‑β signaling is implicated in autoinflammatory phenotypes and interferonopathies, and it contributes to immune remodeling in chronic infection and tumor-associated inflammation, making IFNB1 a widely used axis for mechanistic immunology studies.

IFN-β CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFNB1 expression without altering the underlying DNA sequence.

IFN-β CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFNB1 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 IFNB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFN-β expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFNB1 locus and enabling the study of IFN-β-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFN-β pathway restoration in tumor cells with silenced or reduced IFNB1 expression.

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