p68 RNA Helicase CRISPR Activation Plasmid (h): sc-402401-ACT

DDX5 encodes p68 RNA helicase, a DEAD-box ATP-dependent RNA helicase that remodels RNA–protein complexes to regulate pre-mRNA splicing, ribosome biogenesis, RNA export, and microRNA processing. p68 also functions as a transcriptional co-regulator by interacting with factors such as p53, β-catenin, and ERα, linking RNA metabolism to cell-cycle control and stress-responsive gene expression programs. Through these activities, DDX5 participates in pathways that coordinate RNA processing with chromatin and transcription dynamics, influencing proliferation, differentiation, and DNA damage responses. Dysregulated DDX5 expression or activity has been associated with altered oncogenic signaling and aberrant RNA processing signatures observed across multiple tumor contexts, supporting its use as a mechanistic node in disease-relevant gene regulation studies.

p68 RNA Helicase CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DDX5 expression without altering the underlying DNA sequence.

p68 RNA Helicase CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DDX5 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 DDX5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p68 RNA Helicase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DDX5 locus and enabling the study of p68 RNA Helicase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p68 RNA Helicase pathway restoration in tumor cells with silenced or reduced DDX5 expression.

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