leupaxin CRISPR Activation Plasmid (h): sc-405101-ACT

LPXN encodes leupaxin, a paxillin family adaptor protein enriched at focal adhesions where it coordinates integrin-dependent signaling with actin cytoskeletal remodeling. Through interactions with kinases and scaffold proteins, leupaxin contributes to cell adhesion, migration, and mechanotransduction processes that shape extracellular matrix sensing and downstream transcriptional programs. LPXN activity intersects with focal adhesion and Rho GTPase–linked pathways that regulate cell motility and invasive behavior. Dysregulated expression or signaling involving leupaxin has been reported across multiple tumor contexts and in immune-cell functional states, supporting its use as a mechanistic node in studies of metastasis-associated phenotypes and adhesion-driven signaling.

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

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

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