PARD3A CRISPR Activation Plasmid (m): sc-430042-ACT

Mouse Pard3 encodes the polarity protein PARD3A, a core scaffold of the PAR polarity complex that coordinates apical–basal organization through interactions with aPKC and PAR6. PARD3A helps establish tight junctions and regulates epithelial barrier formation, directed cell migration, and asymmetric cell division by integrating cues from junctional adhesion complexes and cytoskeletal dynamics. Through control of cell–cell contact signaling and polarity-dependent trafficking, PARD3A influences pathways linked to tissue morphogenesis and stem/progenitor cell behavior. Dysregulated PAR complex activity and loss of epithelial polarity are widely associated with invasive phenotypes and progression in multiple disease contexts, making Pard3 a useful node for mechanistic studies of polarity breakdown.

PARD3A CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Pard3 expression without altering the underlying DNA sequence.

PARD3A CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Pard3 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 Pard3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PARD3A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Pard3 locus and enabling the study of PARD3A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PARD3A pathway restoration in tumor cells with silenced or reduced Pard3 expression.

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