WWOX CRISPR Activation Plasmid (h): sc-403070-ACT

WWOX (WW domain-containing oxidoreductase) encodes a tumor suppressor–associated protein mapped to the common fragile site FRA16D and implicated in maintaining genome stability. Human WWOX participates in signaling networks that integrate apoptosis, DNA damage responses, and cellular stress pathways, mediated in part through WW domain interactions with proline-rich partners and modulation of transcriptional regulators. Altered WWOX expression or loss-of-function has been linked to oncogenic transformation and metastatic phenotypes across multiple tissue contexts, and genetic disruption is also associated with neurodevelopmental disorders including epileptic encephalopathy. These properties make WWOX a frequently studied node in pathways controlling proliferation, differentiation, and cell death.

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

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

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