DDX27 CRISPR Activation Plasmid (h): sc-409574-ACT

DDX27 encodes a nucleolar DEAD-box RNA helicase that participates in pre-rRNA processing and ribosome biogenesis, supporting efficient assembly of large ribosomal subunits and global translational capacity. By remodeling RNA–protein complexes in the nucleolus, DDX27 contributes to regulation of cell growth and proliferation programs that depend on tightly controlled protein synthesis. Disruption of ribosome production and nucleolar homeostasis is linked to cellular stress signaling and altered differentiation, making DDX27 relevant for studies of ribosomopathies and cancer-associated translational rewiring. Aberrant DDX27 activity has been associated with proliferative phenotypes and defects in myogenic and developmental processes in model systems, supporting its use as a mechanistic node in RNA metabolism research.

DDX27 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DDX27 expression without altering the underlying DNA sequence.

DDX27 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DDX27 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 DDX27 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDX27 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DDX27 locus and enabling the study of DDX27-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDX27 pathway restoration in tumor cells with silenced or reduced DDX27 expression.

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