The ANAPC7 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-mediated gene disruption product composed of a polyclonal population of A-549 human lung adenocarcinoma epithelial cells harboring targeted mutations in the ANAPC7 locus. This non-clonal format ensures the retention of cellular heterogeneity while providing a robust loss-of-function model, as confirmed by immunoblotting for ANAPC7 protein ablation. The cells are provided as a frozen stock suitable for expansion and downstream assays.
The A-549 cell line is a classic in vitro model for non-small cell lung cancer (NSCLC), derived from a human lung adenocarcinoma and characterized by an oncogenic KRAS (G12S) mutation. These adherent epithelial cells maintain alveolar type II pneumocyte features and are extensively employed in cancer research for studying proliferation, apoptosis, drug metabolism, and metastasis. Their ease of transfection and genetic manipulation makes them particularly suited for CRISPR-based knockout strategies aimed at interrogating mitotic processes in a lung cancer context.
ANAPC7 encodes an essential subunit of the anaphase-promoting complex/cyclosome (APC/C), a multi-subunit E3 ubiquitin ligase that governs mitotic progression. Within the APC/C, ANAPC7 interacts with structural components ANAPC2, ANAPC10, and ANAPC11, and facilitates recruitment of substrate adaptors CDC20 and CDH1. During mitosis, cyclin-dependent kinase 1 (CDK1) and polo-like kinase 1 (PLK1) activate APC/C-CDC20, which polyubiquitinates Securin and Cyclin B1, targeting them for proteasomal degradation to trigger anaphase onset. Spindle assembly checkpoint proteins such as MAD2 inhibit APC/C until chromosomes align. Post-mitotically, APC/C-CDH1 mediates destruction of Aurora A and other mitotic cyclins to maintain G1 stability. Disruption of ANAPC7 compromises APC/C complex integrity, resulting in stabilized Cyclin B and Securin, mitotic arrest, and chromosomal instability.
In the A-549 lung adenocarcinoma context, ANAPC7 knockout is expected to disrupt mitotic progression, leading to aneuploidy and apoptosis. The combination of KRAS-driven proliferation signals with impaired mitotic regulation creates oncogenic stress, providing a platform for exploring tumor-suppressive roles of the APC/C pathway. This model enables dissection of how mitotic catastrophe influences cancer cell fate and response to therapeutic agents, particularly those targeting cell division.
These cells enable a broad spectrum of investigations, including flow cytometric cell-cycle analysis, Western blot detection of Cyclin B1 and Securin accumulation, and co-immunoprecipitation to probe APC/C subunit interactions. Additional applications encompass in vitro ubiquitination assays, MTT/BrdU proliferation assays, Annexin V/PI apoptosis assays, and drug sensitivity profiling with mitotic inhibitors like paclitaxel. Transcriptomic analysis via RNA-seq and chromosome missegregation studies using metaphase spreads further extend utility. For technical support or purchasing information, please contact Ascent Research.
