The CD2AP Knockout NCI-H1299 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human CD2AP gene in a well-characterized non-small cell lung carcinoma background. This loss-of-function model is generated via CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous pool of edited cells that enables robust assessment of CD2AP-dependent phenotypes. The polyclonal format provides a practical tool for investigators studying adaptor protein function without single-cell cloning artifacts, facilitating experiments in cancer cell biology, signal transduction, and cytoskeletal regulation.
NCI-H1299 is a human lung carcinoma cell line with epithelial morphology, originally isolated from a lymph node metastasis of a patient with non-small cell lung cancer. This widely utilized cell line serves as a standard model for lung adenocarcinoma research, particularly in studies of metastatic progression, drug resistance, and oncogenic signaling. NCI-H1299 cells harbor TP53 mutations and exhibit adherent growth, making them amenable to a range of functional assays that probe cell migration, invasion, and cytoskeletal organization. Their metastatic origin renders them a relevant host for investigating genes implicated in tumor cell dissemination.
CD2AP encodes a cytoplasmic adaptor protein that functions as a molecular scaffold, bridging transmembrane receptors to the actin cytoskeleton. It is phosphorylated by Src family kinases and interacts with CD2, Nephrin, Podocalyxin, and Cortactin at the plasma membrane. Downstream, CD2AP recruits and activates WASp and the Arp2/3 complex, driving branched actin polymerization and facilitating receptor-mediated endocytosis, podosome formation, and cell motility. In T-cell receptor signaling, CD2AP couples CD2 engagement to actin remodeling, enabling immune synapse formation. Through its SH3 domains and coiled-coil motifs, CD2AP also binds Synaptojanin and other endocytic regulators, participating in membrane trafficking and clathrin-dependent internalization. These interactions position CD2AP as a critical node linking extracellular cues to dynamic cytoskeletal responses.
In the NCI-H1299 lung cancer model, disruption of CD2AP is predicted to compromise actin-dependent processes that underpin metastatic behavior, including lamellipodial protrusion, matrix degradation, and cell adhesion turnover. Given CD2AP’s role in endocytosis and receptor recycling, its loss may alter the surface expression of integrins or growth factor receptors, potentially modulating sensitivity to targeted therapies. Because CD2AP has been associated with cancer metastasis, this knockout cell population provides a tractable system to dissect the mechanistic contributions of adaptor protein assemblies to non-small cell lung carcinoma aggressiveness and drug resistance, enabling comparative studies with wild-type NCI-H1299 controls.
Researchers can employ the CD2AP Knockout NCI-H1299 Polyclonal Cells in a variety of hypothesis-driven assays. Wound healing and Transwell invasion assays permit quantitative measurement of collective and individual cell migration, respectively, revealing CD2AP-dependent motility. Immunofluorescence staining for F-actin and cortactin can delineate actin filament architecture and podosome or invadopodium formation. Co-immunoprecipitation and Western blotting allow exploration of CD2AP interactomes and downstream signaling events involving WASp, Synaptojanin, and Src kinases. Additionally, cell proliferation and drug sensitivity assays can assess how CD2AP influences chemoresistance in a lung cancer context. For further technical details or to inquire about custom configurations, please contact Ascent Research.
