Zyxin CRISPR Activation Plasmid (h): sc-402295-ACT

Human ZYX encodes zyxin, a focal adhesion LIM-domain scaffold that coordinates actin cytoskeleton remodeling and mechanotransduction at sites of integrin-mediated adhesion. Zyxin interacts with VASP/EVL and α-actinin to support stress fiber maintenance, cell spreading, and directional migration, linking extracellular matrix cues to Rho GTPase-regulated contractility and YAP/TAZ-responsive programs. Through its shuttling between adhesion complexes and the nucleus, zyxin can influence transcriptional responses to mechanical stress and cytoskeletal perturbation. Dysregulated ZYX expression or localization has been associated with altered adhesion signaling and invasive phenotypes in cancer-related models, as well as broader roles in tissue remodeling where cell-matrix dynamics are critical.

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

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

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