paxillin CRISPR Activation Plasmid (m): sc-422546-ACT

Mouse Pxn encodes paxillin, a focal adhesion adaptor that coordinates integrin signaling with actin cytoskeleton remodeling to regulate cell adhesion, spreading, and directional migration. Paxillin scaffolds kinases and GTPase regulators at adhesion sites, linking extracellular matrix cues to pathways such as FAK/Src, PI3K-AKT, and Rho family signaling that control traction forces and mechanotransduction. Through these networks, paxillin influences proliferation, survival, and epithelial–mesenchymal dynamics, making it relevant to studies of invasive cell behavior and tissue remodeling. Dysregulated paxillin signaling and focal adhesion turnover are commonly investigated in models of cancer progression, wound repair, and inflammatory microenvironments.

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

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

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