PARD3B CRISPR Activation Plasmid (h2): sc-406097-ACT-2

Human PARD3B encodes partitioning defective 3 homolog B, a polarity scaffolding protein that localizes to cell–cell junctions and coordinates assembly of apical–basal polarity complexes with partners such as atypical PKC and other PAR proteins. Through PDZ domain–mediated interactions, PARD3B helps regulate tight junction organization, epithelial morphogenesis, and directional migration by integrating cues from cytoskeletal and small GTPase signaling pathways. Altered polarity network function and junctional integrity involving PARD3B has been implicated in processes relevant to tumor progression, invasion, and epithelial barrier dysfunction. Gene editing of PARD3B enables mechanistic studies of polarity-dependent signaling, junction dynamics, and cell-state transitions in human cell models using knockout, knock-in, or domain-specific perturbations.

PARD3B CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous PARD3B expression without altering the underlying DNA sequence.

PARD3B CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PARD3B 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 PARD3B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PARD3B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PARD3B locus and enabling the study of PARD3B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PARD3B pathway restoration in tumor cells with silenced or reduced PARD3B expression.

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