DDX15 CRISPR Activation Plasmid (h): sc-403959-ACT

DHX15 (DDX15) encodes a DEAH-box ATP-dependent RNA helicase that remodels RNA–protein complexes to support pre-mRNA splicing and broader RNA metabolism. It participates in spliceosome dynamics, including intron recognition and catalytic transitions, and has been linked to coordination between transcription and RNA processing. By influencing transcript isoform usage and RNA quality control, DDX15 can modulate gene expression programs that shape cell-cycle progression and stress responses. Dysregulated RNA splicing and helicase activity are frequently associated with oncogenic transformation and hematologic and neurodevelopmental phenotypes, making DHX15 a useful node for mechanistic studies of RNA-processing defects.

DDX15 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DHX15 expression without altering the underlying DNA sequence.

DDX15 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DHX15 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 DHX15 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDX15 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DHX15 locus and enabling the study of DDX15-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDX15 pathway restoration in tumor cells with silenced or reduced DHX15 expression.

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