Radixin CRISPR Activation Plasmid (h): sc-403440-ACT

Human RDX encodes radixin, an ERM family cytoskeletal adaptor that links cortical F-actin to transmembrane proteins to organize membrane domains, regulate cell shape, and stabilize cell–cell and cell–matrix contacts. Radixin participates in remodeling of the actin cortex at structures such as microvilli and adhesion sites and integrates signaling through Rho-family GTPases, PI3K-dependent pathways, and scaffolding complexes that coordinate receptor localization and trafficking. Through its role in mechanotransduction and membrane–cytoskeleton coupling, radixin influences processes including epithelial polarity, migration, and endocytosis. Dysregulated ERM function and altered RDX expression or localization have been associated with phenotypes relevant to cancer cell invasion and metastasis as well as tissue-specific dysfunction in barrier and neuronal contexts.

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

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

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