CDC42 CRISPR Activation Plasmid (h): sc-418353-ACT

CDC42 encodes a ubiquitously expressed Rho-family small GTPase that functions as a molecular switch controlling actin cytoskeleton remodeling, cell polarity, vesicle trafficking, and cell-cycle progression. By cycling between GDP- and GTP-bound states, CDC42 coordinates signaling through effectors such as PAKs, WASP/N-WASP, and the PAR polarity complex, linking receptor cues to cytoskeletal dynamics and transcriptional programs. CDC42 activity intersects with pathways including PI3K–AKT, MAPK, and integrin-mediated adhesion to regulate migration, endocytosis, and junctional integrity. Dysregulated CDC42 signaling has been associated with altered immune cell function, neurodevelopmental abnormalities, and oncogenic phenotypes characterized by aberrant motility and invasion.

CDC42 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDC42 expression without altering the underlying DNA sequence.

CDC42 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDC42 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 CDC42 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CDC42 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDC42 locus and enabling the study of CDC42-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CDC42 pathway restoration in tumor cells with silenced or reduced CDC42 expression.

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