DDX3X CRISPR Activation Plasmid (h): sc-401253-ACT

DDX3X encodes a conserved DEAD-box RNA helicase that remodels RNA secondary structure and ribonucleoprotein complexes to coordinate multiple steps of RNA metabolism, including transcriptional regulation, pre-mRNA processing, mRNA export, translation initiation, and stress granule dynamics. In human cells, DDX3X integrates signaling cues that shape innate immune responses and cellular stress programs, linking RNA sensing to interferon-associated pathways and translational control. Altered DDX3X activity or expression has been associated with dysregulated proliferation, aberrant stress responses, and genome maintenance defects, making it relevant to mechanistic studies of cancer biology and neurodevelopmental disorders. As a multifunctional RNA helicase, DDX3X is frequently used as a node for dissecting post-transcriptional gene regulation and pathway crosstalk.

DDX3X CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DDX3X expression without altering the underlying DNA sequence.

DDX3X CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DDX3X 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 DDX3X transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDX3X expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DDX3X locus and enabling the study of DDX3X-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDX3X pathway restoration in tumor cells with silenced or reduced DDX3X expression.

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