The CCDC88A Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the CCDC88A gene in the near-haploid HAP1 cell line. This heterogeneous pool of HAP1 cells carries targeted gene disruptions at the CCDC88A locus, eliminating functional Girdin protein expression. Because HAP1 cells are haploid, disruption of the single allele provides a complete loss-of-function model, enabling unambiguous characterization of Girdin-dependent processes.
HAP1 cells are a chronic myeloid leukemia-derived, near-haploid, fibroblast-like cell line originally isolated from the KBM-7 line. Their haploid karyotype simplifies knockout studies, as a single allelic disruption yields full loss of gene function. HAP1 cells display adhesion-dependent growth, robust signaling responses, and excellent tractability for imaging, biochemical, and high-throughput assays, making them an ideal platform for studying cytoskeletal dynamics and cell migration.
CCDC88A encodes Girdin, a scaffold protein that integrates PI3K/Akt and Wnt signaling to regulate actin cytoskeleton remodeling and cell motility. Girdin is phosphorylated by Akt downstream of activated PI3K, often triggered by growth factor receptors such as EGFR and IGF1R. Activated Girdin engages Dishevelled (Dvl) and G??i to promote Rac1-dependent actin polymerization, lamellipodia formation, and directed migration. Girdin also sustains mTORC2 signaling and interacts with Par-3 to coordinate cell polarity, thereby linking extracellular cues to the cytoskeleton and facilitating invasive cell behavior.
In the HAP1 background, CCDC88A disruption provides a clean genetic system to dissect Girdin??s functions. The haploid nature ensures that knockout phenotypes are not masked by a wild-type allele, enabling rigorous functional annotation. HAP1 cells exhibit robust growth factor responses and endogenous expression of pathway components, making them well-suited for quantifying migration defects, cytoskeletal reorganization, and alterations in PI3K/Akt or Wnt signal transduction following Girdin loss.
These polyclonal knockout cells are suited for a variety of applications, including transwell migration/invasion assays, wound-healing assays, and immunofluorescence visualization of actin dynamics. Researchers can also perform western blotting to monitor phospho-Akt and downstream effectors, co-immunoprecipitation of Girdin complexes, and transcriptomic profiling to map signaling networks. The cells facilitate drug-target validation studies assessing Girdin??s contribution to kinase inhibitor sensitivity or anti-migratory compound efficacy. For technical support or custom inquiries, please contact Ascent Research.
