Wnt-5b CRISPR Activation Plasmid (h2): sc-402157-ACT-2

Human WNT5B encodes the secreted ligand Wnt-5b, a key mediator of β-catenin–independent (non-canonical) Wnt signaling that regulates cell polarity, directional migration, and cytoskeletal remodeling through pathways such as planar cell polarity and Wnt/Ca²⁺ signaling. Wnt-5b engages Frizzled receptors and co-receptors (including ROR family proteins) to modulate downstream effectors that shape tissue morphogenesis, mesenchymal differentiation, and inflammatory signaling cross-talk. Altered WNT5B expression or signaling has been linked to dysregulated developmental programs and is frequently investigated in contexts such as tumor cell invasiveness, metabolic and adipogenic regulation, and fibrotic remodeling. Gene editing of WNT5B supports mechanistic studies of ligand-specific Wnt network wiring, receptor usage, and transcriptional and phenotypic outcomes in engineered cell models and organoid systems.

Wnt-5b CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous WNT5B expression without altering the underlying DNA sequence.

Wnt-5b CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the WNT5B 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 WNT5B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Wnt-5b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native WNT5B locus and enabling the study of Wnt-5b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Wnt-5b pathway restoration in tumor cells with silenced or reduced WNT5B expression.

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