Wnt-7b CRISPR Activation Plasmid (m): sc-423722-ACT

Wnt7b encodes the secreted glycoprotein Wnt-7b, a key ligand in canonical and non-canonical Wnt signaling that regulates cell fate specification, proliferation, and tissue patterning during development. In mouse, Wnt-7b activity contributes to organogenesis and epithelial–mesenchymal signaling, influencing processes such as angiogenesis, branching morphogenesis, and bone remodeling via β-catenin–dependent transcriptional programs. Dysregulated Wnt-7b/Wnt pathway signaling is associated with aberrant differentiation and remodeling phenotypes relevant to developmental disorders, fibrosis-associated tissue changes, and cancer biology. As a pathway cue, Wnt-7b is frequently used to interrogate context-specific transcriptional networks and morphogen gradients in vivo and in cultured cell systems.

Wnt-7b CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Wnt7b expression without altering the underlying DNA sequence.

Wnt-7b CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Wnt7b 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 Wnt7b transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Wnt-7b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Wnt7b locus and enabling the study of Wnt-7b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Wnt-7b pathway restoration in tumor cells with silenced or reduced Wnt7b expression.

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