DDX60 CRISPR Activation Plasmid (m): sc-433350-ACT

Mouse Ddx60 encodes DDX60, an interferon-inducible DExD/H-box RNA helicase that functions as a cytosolic sensor and regulator of antiviral innate immunity. DDX60 participates in RNA metabolism and pattern-recognition pathways by promoting RIG-I/MDA5-dependent signaling, amplifying type I interferon responses, and supporting ISG expression programs that restrict viral replication. Through interactions with RNA substrates and immune signaling components, DDX60 influences NF-κB and IRF-driven transcriptional networks and shapes cellular responses to double-stranded RNA stress. Dysregulation of Ddx60 activity is therefore relevant to studies of inflammatory signaling, host–pathogen interactions, and immune-associated disease mechanisms in mouse models.

DDX60 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ddx60 expression without altering the underlying DNA sequence.

DDX60 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ddx60 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 Ddx60 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDX60 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ddx60 locus and enabling the study of DDX60-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDX60 pathway restoration in tumor cells with silenced or reduced Ddx60 expression.

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