IFIT1 CRISPR Activation Plasmid (h): sc-403066-ACT

Interferon induced protein with tetratricopeptide repeats 1 (IFIT1) is an interferon-stimulated gene product that binds viral and aberrantly capped RNAs, including 5′-triphosphate and cap0 structures, to restrict translation and amplify innate immune defenses. IFIT1 functions within type I/III interferon signaling downstream of JAK–STAT activation, integrating with pattern-recognition receptor pathways such as RIG-I/MDA5 and TLR3 to shape the antiviral state. Through interactions with translation initiation machinery and other IFIT family members, it modulates RNA metabolism and protein synthesis programs during immune activation. Dysregulated IFIT1 expression has been associated with altered antiviral responses and inflammatory transcriptional signatures observed in infection, interferonopathies, and tumor-immune microenvironments, making it a useful marker and mechanistic node for immunology and virology research.

IFIT1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFIT1 expression without altering the underlying DNA sequence.

IFIT1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFIT1 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 IFIT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFIT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFIT1 locus and enabling the study of IFIT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFIT1 pathway restoration in tumor cells with silenced or reduced IFIT1 expression.

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