The CCDC25 Knockout HAP1 Polyclonal Cells constitute a versatile CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the CCDC25 gene. This product is generated in the HAP1 host cell line using a polyclonal pool of cells, providing a heterogeneous loss-of-function model that avoids clonal artifacts. The polyclonal format is particularly suited to studies where population-level functional effects of CCDC25 ablation are assessed without the bias of single-cell-derived clones, enabling robust and reproducible analyses of receptor-mediated signaling and cell migration. Researchers can employ these cells to interrogate the role of the CCDC25 receptor in the context of neutrophil extracellular trap (NET)-driven processes, with broad utility in cancer biology and inflammation research.
HAP1 is a near-haploid, fibroblast-like cell line originally derived from the KBM-7 chronic myeloid leukemia (CML) cell line, which was established from a patient in blast crisis. As an adherent cell type with a partially haploid karyotype, HAP1 serves as a highly tractable genetic screening platform, facilitating gene knockout, mutagenesis, and functional genomics studies. Its derivation from a CML background imparts unique signaling characteristics related to hematopoietic malignancy, yet its fibroblast-like morphology and adhesive properties render it amenable to standard migration assays, immunofluorescence, and biochemical analyses. This host cell context is particularly relevant for modeling mechanisms of metastasis and cell motility, given that CML cells can exhibit invasive phenotypes, and the HAP1 line retains key integrin and actin remodeling pathways.
The CCDC25 gene encodes a transmembrane receptor that specifically recognizes DNA within neutrophil extracellular traps (NETs), a form of extracellular DNA released by neutrophils during inflammation. Upon NET-DNA binding, CCDC25 activates an intracellular signaling cascade involving integrin-linked kinase (ILK) and parvin, which subsequently triggers the small GTPases Rac1 and Cdc42 to drive localized actin polymerization and cell migration. This ILK-parvin pathway is central to CCDC25-mediated enhancement of metastasis, as demonstrated in models of breast cancer progression. Upstream, the signaling is initiated by NET-derived DNA, while downstream effectors include ILK, parvin, Rac1, Cdc42, and F-actin remodeling. Interacting factors such as extracellular DNA and the ILK-parvin complex directly associate with the cytoplasmic domain of CCDC25, transducing the external NET signal into cytoskeletal reorganization.
In HAP1 cells, knockout of CCDC25 eliminates the receptor for NET-DNA, thereby blocking ILK-parvin pathway activation and downstream actin remodeling. This loss-of-function model effectively inhibits NET-induced cell migration, mirroring the ablation of a key metastatic signaling axis. Given the HAP1 cell line??s CML origin and its established use in genetic screens, the CCDC25 knockout polyclonal population provides a unique tool to dissect the intersection of leukemogenic backgrounds and innate immune-derived prometastatic cues. Researchers can investigate whether CML-related kinase dependencies or integrin profiles modulate CCDC25 signaling, offering insights into how hematopoietic malignancies might exploit NET-mediated pathways for disease progression.
These cells support a wide array of experimental applications, including detailed CCDC25 functional studies, NET-mediated metastasis research, and validation of potential drug targets. For example, NET-induced migration assays and transwell migration setups can be employed to quantify the loss of chemotactic and invasive capacity following CCDC25 ablation. Western blotting, qRT-PCR, and flow cytometry enable verification of pathway disruption at the protein and transcript levels, while immunofluorescence for F-actin reveals altered actin dynamics. Additionally, the polyclonal knockout population is well-suited for screening inhibitors of the CCDC25-ILK interaction, facilitating early-stage drug discovery in oncology. For further information, contact Ascent Research.
