DDX60L CRISPR Activation Plasmid (h): sc-413888-ACT

DDX60L (DEAD-box helicase 60-like) is a human RNA helicase implicated in innate antiviral defense, functioning in cytosolic RNA sensing and interferon-stimulated gene programs. It is associated with regulation of RNA metabolism and RIG-I–like receptor signaling pathways that shape type I interferon responses and downstream transcriptional networks. As an interferon-inducible factor, DDX60L is frequently studied in the context of host–pathogen interactions and inflammatory signaling. Dysregulated interferon and RNA-sensing pathways are relevant to infection biology and immune-mediated disease mechanisms, supporting investigation of DDX60L-dependent control of viral replication and cytokine responses.

DDX60L CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DDX60L expression without altering the underlying DNA sequence.

DDX60L CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DDX60L 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 DDX60L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDX60L expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DDX60L locus and enabling the study of DDX60L-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDX60L pathway restoration in tumor cells with silenced or reduced DDX60L expression.

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