Plectin CRISPR Activation Plasmid (h): sc-401494-ACT

Human PLEC encodes plectin, a large cytolinker protein that integrates intermediate filaments with actin microfilaments and microtubules to stabilize cell architecture under mechanical stress. By anchoring cytoskeletal networks to membrane-associated complexes such as hemidesmosomes and focal adhesions, plectin supports cell–matrix adhesion, mechanotransduction, and dynamic remodeling during migration and differentiation. Plectin-dependent organization of filament systems influences signaling nodes linked to cellular integrity and stress responses, including pathways governing epithelial resilience and myofiber stability. Altered PLEC function or expression has been associated with tissue fragility phenotypes and neuromuscular and epithelial pathologies, making it relevant for studies of cytoskeletal cross-talk and barrier maintenance.

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

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

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