DDX36 CRISPR Activation Plasmid (h): sc-404744-ACT

DHX36 encodes the human RNA/DNA helicase DDX36, a DEAH-box helicase that preferentially binds and resolves G-quadruplex structures in both RNA and DNA. By unwinding these secondary structures, DDX36 helps regulate transcriptional and post-transcriptional programs including mRNA turnover, translation, and maintenance of genome stability. The protein has been linked to control of cell proliferation and stress-responsive gene expression, processes frequently perturbed in cancer and other diseases where G-quadruplex-mediated regulation is altered. DDX36 function is also relevant to pathways governing RNA metabolism and nucleic acid surveillance that shape oncogenic signaling and cellular differentiation states.

DDX36 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DHX36 expression without altering the underlying DNA sequence.

DDX36 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DHX36 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 DHX36 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDX36 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DHX36 locus and enabling the study of DDX36-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDX36 pathway restoration in tumor cells with silenced or reduced DHX36 expression.

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