IFIT3 CRISPR Activation Plasmid (h): sc-403557-ACT

Interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) is an interferon-stimulated gene product that amplifies innate antiviral defenses downstream of type I interferon and pattern-recognition receptor signaling. IFIT3 functions within cytoplasmic IFIT complexes to modulate translation and RNA sensing, influencing ISG induction, JAK–STAT signaling output, and inflammatory transcriptional programs. By shaping the magnitude and persistence of interferon responses, IFIT3 is frequently studied in the context of host–pathogen interactions, immune regulation, and interferon-driven cellular stress. Dysregulated IFIT3 expression has been associated with altered immune microenvironments and inflammatory states, supporting its relevance in mechanistic studies of infection biology and immune-associated disease processes.

IFIT3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFIT3 expression without altering the underlying DNA sequence.

IFIT3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFIT3 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 IFIT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFIT3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFIT3 locus and enabling the study of IFIT3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFIT3 pathway restoration in tumor cells with silenced or reduced IFIT3 expression.

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