ANAPC16 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of human A-549 lung adenocarcinoma epithelial cells with targeted disruption of the ANAPC16 gene. This loss-of-function model enables investigation of anaphase-promoting complex/cyclosome (APC/C) E3 ubiquitin ligase function in a KRAS-mutant lung cancer background. The polyclonal format preserves cell heterogeneity while ensuring robust knockout representation across the population, making it suitable for diverse experimental assays.
A-549 cells are an adherent epithelial cell line derived from human lung adenocarcinoma, widely used as a model of alveolar type II epithelium. They harbor an activating KRAS mutation, rendering them particularly relevant for studying oncogenic signaling and chromosomal instability in non-small cell lung cancer. Their well-characterized growth characteristics and responsiveness to mitotic perturbations make them an ideal host for dissecting cell cycle regulation and APC/C-dependent processes.
ANAPC16 encodes a constitutive subunit of the APC/C complex, a multi-subunit E3 ubiquitin ligase that governs mitotic progression by targeting key substrates for proteasomal degradation. The APC/C is activated by co-activators CDC20 and CDH1, which are in turn regulated by CDK1 phosphorylation and checkpoint proteins such as MAD2L1 and BUB1B. ANAPC16 interacts with other core APC/C subunits including ANAPC2, ANAPC10, ANAPC11, and CDC26 to maintain complex integrity. Downstream substrates of APC/C activity include cyclin B1 and securin, whose degradation triggers the metaphase-to-anaphase transition, along with mitotic kinases PLK1 and Aurora A, which govern spindle assembly and cytokinesis. Disruption of ANAPC16 can impair APC/C assembly or activity, leading to mitotic arrest, aneuploidy, and chromosomal instability??hallmarks of aggressive cancers.
In the KRAS-driven A-549 background, loss of ANAPC16 provides a powerful system to dissect APC/C-dependent tumor-suppressive mechanisms and the consequences of mitotic stress in lung adenocarcinoma. Combined with the intrinsic genomic instability of the KRAS-mutant line, ANAPC16 knockout may exacerbate spindle assembly checkpoint defects and reveal synthetic lethal vulnerabilities. This model thus permits exploration of the interplay between oncogenic signaling, ubiquitin-mediated proteolysis, and cell cycle fidelity.
Researchers can employ these polyclonal knockout cells for western blotting of cell cycle markers (e.g., cyclin B1, securin), flow cytometric cell cycle analysis, and live-cell imaging to monitor mitotic progression and spindle defects. Co-immunoprecipitation assays enable assessment of APC/C complex assembly, while ubiquitination assays probe the activity of the ligase toward its substrates. Viability assays under mitotic stress (e.g., spindle poisons) can identify potential therapeutic targets. This product is suited for studies in cancer biology, chromosomal instability, aneuploidy, and drug target validation. For further technical details, please contact Ascent Research.
