girdin CRISPR Activation Plasmid (h): sc-402236-ACT

Human CCDC88A encodes girdin, a large actin-binding scaffold that integrates growth factor and GPCR signaling with cytoskeletal remodeling to coordinate cell polarity, directed migration, and adhesion. Girdin functions downstream of PI3K–AKT and interacts with heterotrimeric G proteins and polarity regulators to influence lamellipodia formation, vesicle trafficking, and receptor signaling dynamics. Through these roles, CCDC88A contributes to processes such as neurite outgrowth and epithelial organization, and altered expression or signaling coupling has been associated with aberrant motility and survival phenotypes observed in cancer and other proliferative or neurodevelopmental disease contexts. Its pathway connectivity makes CCDC88A a useful node for dissecting how cytoskeletal architecture modulates signal transduction and cell-state transitions.

girdin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCDC88A expression without altering the underlying DNA sequence.

girdin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCDC88A 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 CCDC88A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous girdin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCDC88A locus and enabling the study of girdin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of girdin pathway restoration in tumor cells with silenced or reduced CCDC88A expression.

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