IRF-7 CRISPR Activation Plasmid (m): sc-424785-ACT

Mouse Irf7 encodes IRF-7, a master transcriptional regulator of type I interferon responses that coordinates antiviral innate immunity downstream of pattern recognition receptors such as RIG-I–like receptors and Toll-like receptors. Upon activation, IRF-7 integrates signaling from TBK1/IKKε-dependent phosphorylation and nuclear translocation to drive interferon-stimulated gene programs through the JAK–STAT axis. Irf7 activity shapes inflammatory signaling, dendritic cell and macrophage responses, and cross-talk with NF-κB–mediated cytokine networks. Dysregulated IRF-7–dependent interferon signatures are relevant to models of viral susceptibility, chronic inflammation, and autoimmune-like pathology, supporting mechanistic studies of interferon-driven disease processes.

IRF-7 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Irf7 expression without altering the underlying DNA sequence.

IRF-7 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Irf7 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 Irf7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IRF-7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Irf7 locus and enabling the study of IRF-7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IRF-7 pathway restoration in tumor cells with silenced or reduced Irf7 expression.

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