DDX30 CRISPR Activation Plasmid (h): sc-411632-ACT

DHX30 (DDX30) encodes an ATP-dependent DEAD-box RNA helicase that modulates RNA secondary structure to support RNA metabolism, including aspects of transcriptional regulation, RNA processing, and translation. As a helicase, DDX30 contributes to ribonucleoprotein remodeling and RNA quality-control processes that influence cellular stress responses and mitochondrial and cytosolic gene expression programs. Altered RNA helicase activity can perturb proteostasis and innate immune signaling networks through effects on RNA sensing and transcript fate. Dysregulation of DHX30 has been associated with neurodevelopmental phenotypes and broader disease-relevant transcriptomic changes, making it a useful node for studying RNA-centric mechanisms in human cells.

DDX30 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DHX30 expression without altering the underlying DNA sequence.

DDX30 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DHX30 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 DHX30 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDX30 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DHX30 locus and enabling the study of DDX30-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDX30 pathway restoration in tumor cells with silenced or reduced DHX30 expression.

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