Wnt-11 CRISPR Activation Plasmid (h): sc-402655-ACT

Human WNT11 encodes the secreted glycoprotein Wnt-11, a non-canonical Wnt ligand that primarily signals through planar cell polarity and Wnt/Ca2+ pathways to coordinate cell polarity, directional migration, and tissue morphogenesis. Wnt-11 modulates cytoskeletal dynamics and adhesion programs via downstream effectors such as Rho family GTPases and JNK, and it can contextually intersect with β-catenin–dependent transcription. During development and in adult tissues, WNT11 contributes to organogenesis and maintenance of epithelial and mesenchymal states. Dysregulated WNT11 signaling has been associated with altered migratory phenotypes and remodeling programs observed across multiple disease contexts, supporting its use as a mechanistic node in pathway and phenotype studies.

Wnt-11 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous WNT11 expression without altering the underlying DNA sequence.

Wnt-11 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the WNT11 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 WNT11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Wnt-11 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native WNT11 locus and enabling the study of Wnt-11-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Wnt-11 pathway restoration in tumor cells with silenced or reduced WNT11 expression.

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