Dkk-4 CRISPR Activation Plasmid (h): sc-406386-ACT

Human DKK4 encodes Dickkopf-related protein 4 (Dkk-4), a secreted modulator of Wnt signaling that influences β-catenin–dependent transcription and downstream programs controlling proliferation, differentiation, and epithelial tissue organization. By engaging Wnt co-receptors and tuning pathway output, Dkk-4 can shift cell fate decisions and remodeling responses in developmental and adult contexts. Dysregulated DKK4 expression has been associated with altered Wnt pathway activity in multiple disease-relevant settings, including tumor biology and tissue fibrosis, where changes in signaling balance affect invasion, stem-like phenotypes, and microenvironmental interactions. As a pathway node, DKK4 is frequently studied for its role in cross-talk with growth factor signaling and regulation of extracellular signaling gradients.

Dkk-4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DKK4 expression without altering the underlying DNA sequence.

Dkk-4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DKK4 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 DKK4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dkk-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DKK4 locus and enabling the study of Dkk-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dkk-4 pathway restoration in tumor cells with silenced or reduced DKK4 expression.

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