Wnt-4 CRISPR Activation Plasmid (m): sc-423717-ACT

Mouse Wnt4 encodes Wnt-4, a secreted glycoprotein ligand that regulates cell fate decisions, morphogenesis, and tissue patterning through WNT signaling. Wnt-4 can engage Frizzled/LRP receptor complexes to modulate canonical β-catenin–dependent transcription as well as non-canonical pathways influencing polarity, migration, and cytoskeletal dynamics. In development and adult tissue homeostasis, Wnt-4 contributes to organogenesis and differentiation programs, with dysregulated WNT4 signaling implicated in congenital urogenital phenotypes, aberrant epithelial–stromal interactions, and oncogenic pathway rewiring in multiple contexts. These features make Wnt4 a useful node for studying pathway crosstalk with TGF-β, Notch, and MAPK signaling and for dissecting transcriptional programs downstream of WNT activation.

Wnt-4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Wnt4 expression without altering the underlying DNA sequence.

Wnt-4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Wnt4 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 Wnt4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Wnt-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Wnt4 locus and enabling the study of Wnt-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Wnt-4 pathway restoration in tumor cells with silenced or reduced Wnt4 expression.

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