Cdc42EP4 CRISPR Activation Plasmid (h): sc-408523-ACT

CDC42EP4 encodes Cdc42EP4, a Cdc42/Rho GTPase effector that binds septins and actin-associated machinery to coordinate cytoskeletal remodeling. It contributes to stress fiber formation, focal adhesion dynamics, and changes in cell shape that influence migration and adhesion through Rho GTPase–dependent signaling networks. By modulating actomyosin organization and cell polarity programs, Cdc42EP4 is relevant to studies of epithelial–mesenchymal–like transitions, invasive behavior, and microenvironmental responses in human cells. Altered CDC42EP4 expression has been reported in contexts of dysregulated motility and cytoskeletal control, supporting its investigation in cancer biology, fibrosis-associated remodeling, and developmental cell movement models.

Cdc42EP4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDC42EP4 expression without altering the underlying DNA sequence.

Cdc42EP4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDC42EP4 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 CDC42EP4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cdc42EP4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDC42EP4 locus and enabling the study of Cdc42EP4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cdc42EP4 pathway restoration in tumor cells with silenced or reduced CDC42EP4 expression.

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