IRF-1 CRISPR Activation Plasmid (h): sc-400551-ACT

Human IRF1 encodes interferon regulatory factor 1 (IRF-1), a DNA-binding transcription factor that integrates interferon and inflammatory cues to control antiviral defense, antigen presentation, and cell fate decisions. IRF-1 functions downstream of JAK/STAT signaling and cooperates with NF-κB and IRF family members to regulate transcription of interferon-stimulated genes, cytokine pathways, and components of MHC class I processing. Through regulation of immune surveillance, apoptosis, and growth control programs, IRF1 activity is frequently studied in contexts of infection biology, tumor–immune interactions, and chronic inflammatory states. Dysregulated IRF-1 signaling has been associated with altered innate and adaptive immune responses and changes in cellular sensitivity to stress and cytokine stimulation.

IRF-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IRF1 expression without altering the underlying DNA sequence.

IRF-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IRF1 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 IRF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IRF-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IRF1 locus and enabling the study of IRF-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IRF-1 pathway restoration in tumor cells with silenced or reduced IRF1 expression.

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