IFI-44L CRISPR Activation Plasmid (h): sc-406752-ACT

IFI44L (interferon induced protein 44 like) encodes IFI-44L, an interferon-stimulated gene product that is rapidly induced downstream of type I interferon and JAK/STAT signaling during innate antiviral responses. IFI-44L expression is commonly used as a molecular readout of interferon pathway engagement and is linked to transcriptional programs that shape cellular responses to viral infection and inflammatory cues. Dysregulated IFI44L expression and promoter methylation have been reported across interferon-associated conditions, including systemic autoimmune disease signatures and infection-related immune activation states. These properties make IFI44L a useful target for studying interferon-driven gene networks, immune cell activation, and host–pathogen response biology.

IFI-44L CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFI44L expression without altering the underlying DNA sequence.

IFI-44L CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFI44L 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 IFI44L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFI-44L expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFI44L locus and enabling the study of IFI-44L-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFI-44L pathway restoration in tumor cells with silenced or reduced IFI44L expression.

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