Prickle4 CRISPR Activation Plasmid (h): sc-412059-ACT

PRICKLE4 encodes Prickle4, a member of the Prickle family of planar cell polarity regulators that coordinate directional cell behaviors during development and tissue organization. Prickle proteins participate in non-canonical Wnt/PCP signaling, influencing cytoskeletal dynamics, cell polarity establishment, and polarized migration, with downstream effects on morphogenesis and epithelial architecture. Dysregulation of PCP components can perturb cell movement and polarity-dependent processes, which are frequently implicated in developmental abnormalities and in the altered migratory programs observed in cancer biology. PRICKLE4 expression and pathway activity are therefore of interest for mechanistic studies linking Wnt/PCP signaling to cell shape control, adhesion, and tissue patterning.

Prickle4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRICKLE4 expression without altering the underlying DNA sequence.

Prickle4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRICKLE4 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 PRICKLE4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Prickle4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRICKLE4 locus and enabling the study of Prickle4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Prickle4 pathway restoration in tumor cells with silenced or reduced PRICKLE4 expression.

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