Wnt-8b CRISPR Activation Plasmid (h): sc-405077-ACT

WNT8B encodes the secreted glycoprotein Wnt-8b, a ligand in the Wnt signaling network that regulates cell fate specification, proliferation, and tissue patterning. Wnt-8b primarily influences canonical Wnt/β-catenin transcriptional programs by engaging Frizzled and LRP co-receptors, with downstream effects on developmental gene expression and differentiation trajectories. In human biology, WNT8B activity is closely linked to neurodevelopmental processes and regional patterning in the central nervous system, and altered Wnt pathway tone is frequently studied in the context of oncogenic signaling and aberrant growth control. Modulating WNT8B expression is therefore relevant for dissecting pathway cross-talk that includes TCF/LEF-mediated transcription, stem-like cell states, and context-dependent differentiation outcomes.

Wnt-8b CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous WNT8B expression without altering the underlying DNA sequence.

Wnt-8b CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the WNT8B 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 WNT8B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Wnt-8b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native WNT8B locus and enabling the study of Wnt-8b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Wnt-8b pathway restoration in tumor cells with silenced or reduced WNT8B expression.

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