IFI-16 CRISPR Activation Plasmid (h): sc-416568-ACT

Human IFI16 (IFI-16) is a nuclear DNA sensor and interferon-inducible protein that helps coordinate innate immune surveillance of aberrant or foreign nucleic acids. IFI-16 participates in STING-dependent interferon signaling and inflammasome-associated responses, influencing transcriptional programs linked to antiviral defense, cell-cycle control, and chromatin-associated regulation. Through interactions with DNA damage and transcriptional machinery, IFI-16 can modulate cellular stress responses and senescence-associated pathways. Dysregulated IFI16 activity has been associated with inflammatory signaling, viral restriction phenotypes, and immune-related disease mechanisms, making it a useful node for studying nucleic-acid sensing and interferon pathway crosstalk.

IFI-16 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFI16 expression without altering the underlying DNA sequence.

IFI-16 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFI16 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 IFI16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFI-16 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFI16 locus and enabling the study of IFI-16-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFI-16 pathway restoration in tumor cells with silenced or reduced IFI16 expression.

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