frizzled-3 CRISPR Activation Plasmid (h): sc-403362-ACT

FZD3 encodes frizzled-3, a seven-transmembrane Wnt receptor that transduces extracellular Wnt cues into intracellular signaling programs that regulate cell polarity, migration, and differentiation. As a core component of Wnt pathways, frizzled-3 can influence β-catenin–dependent transcription as well as non-canonical planar cell polarity and calcium-associated signaling, depending on ligand and cellular context. FZD3 activity is tightly linked to neurodevelopmental processes such as axon guidance and synaptic organization, and dysregulation of Wnt–FZD signaling has been associated with aberrant proliferation and invasive phenotypes in cancer-relevant models. These properties make frizzled-3 a useful node for interrogating Wnt network wiring, receptor–ligand specificity, and pathway cross-talk in human cells.

frizzled-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FZD3 expression without altering the underlying DNA sequence.

frizzled-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FZD3 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 FZD3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous frizzled-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FZD3 locus and enabling the study of frizzled-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of frizzled-3 pathway restoration in tumor cells with silenced or reduced FZD3 expression.

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