beta-catenin CRISPR Activation Plasmid (m): sc-419477-ACT

Ctnnb1 encodes beta-catenin, a multifunctional armadillo-repeat protein that links cadherin-based adherens junctions to the actin cytoskeleton and serves as a central transcriptional co-activator in canonical Wnt signaling. Upon Wnt pathway activation, stabilized beta-catenin accumulates in the nucleus and partners with TCF/LEF factors to regulate gene programs controlling proliferation, differentiation, epithelial–mesenchymal dynamics, and stem cell maintenance. Tight regulation of beta-catenin turnover by the APC/AXIN/GSK3β destruction complex is essential for tissue homeostasis, and disruption of this axis is strongly associated with oncogenic signaling. In mouse models, Ctnnb1 perturbation is widely used to interrogate development, regeneration, and disease-relevant signaling networks across intestine, liver, skin, bone, and immune compartments.

beta-catenin CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ctnnb1 expression without altering the underlying DNA sequence.

beta-catenin CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ctnnb1 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 Ctnnb1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous beta-catenin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ctnnb1 locus and enabling the study of beta-catenin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of beta-catenin pathway restoration in tumor cells with silenced or reduced Ctnnb1 expression.

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