NDH II CRISPR Activation Plasmid (h): sc-402757-ACT

Human DHX9 encodes the DExH-box helicase NDH II, a multifunctional RNA/DNA helicase that regulates transcription, pre-mRNA processing, mRNA export, translation, and maintenance of genome stability. NDH II participates in R-loop metabolism and resolves structured nucleic acids, linking it to DNA damage response signaling and replication stress pathways. Through interactions with ribonucleoprotein complexes and promoter-associated nucleic acids, DHX9 can influence innate immune and stress-responsive gene expression programs. Altered DHX9 activity or expression has been associated with dysregulated RNA processing and chromatin-linked instability observed across multiple disease-relevant cellular contexts, supporting its use as a mechanistic target in functional genomics.

NDH II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DHX9 expression without altering the underlying DNA sequence.

NDH II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DHX9 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 DHX9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NDH II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DHX9 locus and enabling the study of NDH II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NDH II pathway restoration in tumor cells with silenced or reduced DHX9 expression.

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