DDX17 CRISPR Activation Plasmid (h): sc-401338-ACT

DDX17 (DEAD-box helicase 17) is a human ATP-dependent RNA helicase that remodels RNA and ribonucleoprotein complexes to coordinate multiple steps of RNA metabolism. It participates in pre-mRNA splicing, transcriptional co-regulation, microRNA biogenesis, and ribosome-related RNA processing, linking RNA helicase activity to gene expression programs that govern proliferation and differentiation. DDX17 function intersects with signaling-dependent transcriptional networks and chromatin-associated regulatory complexes, shaping context-specific transcript isoforms and RNA stability. Dysregulation of DDX17-associated RNA processing and transcriptional control has been implicated in oncogenic and developmental pathways, making it relevant for mechanistic studies of altered gene expression in disease models.

DDX17 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DDX17 expression without altering the underlying DNA sequence.

DDX17 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DDX17 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 DDX17 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDX17 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DDX17 locus and enabling the study of DDX17-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDX17 pathway restoration in tumor cells with silenced or reduced DDX17 expression.

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