Wnt-10a CRISPR Activation Plasmid (h): sc-403285-ACT

WNT10A encodes Wnt-10a, a secreted Wnt family ligand that regulates cell fate specification, proliferation, and tissue patterning through canonical β-catenin/TCF signaling and context-dependent noncanonical Wnt pathways. Wnt-10a activity influences epithelial–mesenchymal interactions and differentiation programs, with prominent roles in development and maintenance of ectodermal appendages such as hair follicles and teeth. Dysregulated WNT10A signaling has been linked to congenital ectodermal dysplasias and odontogenic defects, and altered expression is also studied in cancers where Wnt pathway remodeling affects tumor cell behavior and stromal responses. As a pathway node, WNT10A is frequently used to interrogate Wnt ligand-specific effects on transcriptional networks, morphogenesis, and stem/progenitor cell dynamics.

Wnt-10a CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous WNT10A expression without altering the underlying DNA sequence.

Wnt-10a CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the WNT10A 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 WNT10A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Wnt-10a expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native WNT10A locus and enabling the study of Wnt-10a-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Wnt-10a pathway restoration in tumor cells with silenced or reduced WNT10A expression.

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