PARD6B CRISPR Activation Plasmid (h): sc-401699-ACT

Human PARD6B encodes Par-6 beta, a PDZ-domain polarity scaffold that assembles the PAR polarity complex with PAR3 and atypical PKC to establish apical–basal polarity and coordinate tight junction formation. Through interactions with small GTPases such as CDC42 and RAC1, PARD6B helps regulate cytoskeletal dynamics, oriented cell division, and epithelial morphogenesis, with downstream effects on migration and tissue architecture. Altered polarity signaling involving PARD6B has been associated with epithelial disorganization and invasive phenotypes in cancer-relevant contexts, and it is also studied in developmental processes and barrier function. As a signaling hub at cell–cell junctions, PARD6B provides a tractable entry point for dissecting polarity-dependent transcriptional and phenotypic programs.

PARD6B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PARD6B expression without altering the underlying DNA sequence.

PARD6B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PARD6B 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 PARD6B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PARD6B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PARD6B locus and enabling the study of PARD6B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PARD6B pathway restoration in tumor cells with silenced or reduced PARD6B expression.

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