Pinin CRISPR Activation Plasmid (h): sc-405373-ACT

Human PNN encodes pinin, a nuclear and desmosome-associated protein that links epithelial cell–cell adhesion with transcriptional and RNA-processing programs. Pinin participates in mRNA splicing and exon junction complex–related processes, influencing alternative splicing outcomes and gene expression networks that shape epithelial differentiation and tissue integrity. Through its roles in desmosomal organization, nuclear speckles, and regulation of transcript maturation, PNN can modulate pathways involved in cell polarity, migration, and stress-responsive gene regulation. Altered PNN expression or localization has been associated with changes in epithelial phenotype and has been investigated in the context of tumor progression, metastasis-related transcriptional programs, and other disorders featuring adhesion and splicing dysregulation.

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

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

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