Pygopus 2 CRISPR Activation Plasmid (h): sc-402894-ACT

PYGO2 encodes Pygopus 2, a nuclear coactivator that functions as a chromatin-associated effector of canonical Wnt/β-catenin signaling. By engaging β-catenin/TCF transcriptional complexes and supporting recruitment of additional transcriptional machinery, PYGO2 contributes to regulation of proliferation, lineage specification, and stem-like transcriptional programs. Pygopus 2 activity is linked to epigenetic control of Wnt target loci and intersects with processes such as cell-cycle progression and epithelial differentiation. Dysregulated PYGO2 expression and Wnt pathway output have been associated with oncogenic transcriptional states and tumor biology in multiple contexts, supporting its relevance as a mechanistic node for pathway-focused studies.

Pygopus 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PYGO2 expression without altering the underlying DNA sequence.

Pygopus 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PYGO2 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 PYGO2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pygopus 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PYGO2 locus and enabling the study of Pygopus 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pygopus 2 pathway restoration in tumor cells with silenced or reduced PYGO2 expression.

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