Wnt-10b CRISPR Activation Plasmid (m2): sc-423710-ACT-2

Mouse Wnt10b encodes the secreted glycoprotein Wnt-10b, a canonical Wnt ligand that activates Frizzled/LRP receptor complexes to promote β-catenin stabilization and TCF/LEF-dependent transcription. Wnt-10b regulates embryonic patterning and adult tissue homeostasis by controlling cell fate decisions, proliferation, and differentiation, with well-established roles in osteoblastogenesis, adipogenesis suppression, and hair follicle and mammary gland biology. Dysregulated Wnt10b signaling has been linked to altered bone mass, metabolic and adipose phenotypes, and context-dependent tumor-related Wnt pathway activation, making it a useful target for mechanistic studies of Wnt/β-catenin dynamics. Gene editing of Wnt10b in mouse cells or models enables functional dissection of ligand-specific Wnt signaling, pathway crosstalk, and developmental or metabolic phenotypes using reporter assays, differentiation systems, and in vivo lineage analyses.

Wnt-10b CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Wnt10b expression without altering the underlying DNA sequence.

Wnt-10b CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Wnt10b 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 Wnt10b transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Wnt-10b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Wnt10b locus and enabling the study of Wnt-10b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Wnt-10b pathway restoration in tumor cells with silenced or reduced Wnt10b expression.

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