The ANKRD13A Knockout A-549 Polyclonal Cells product comprises a heterogeneous population of A-549 lung adenocarcinoma cells in which the ANKRD13A locus has been disrupted via CRISPR/Cas9. This polyclonal knockout pool is generated without single-cell cloning, maintaining the genetic diversity of the parental cells while achieving efficient target-gene ablation. The resulting loss of ANKRD13A protein provides a rigorous loss-of-function model for dissecting its cellular functions.
Derived from a 58-year-old male lung adenocarcinoma patient, the A-549 cell line is an epithelial model for non-small cell lung cancer. These cells harbor active oncogenic signaling through EGFR and other RTKs and depend on the endo-lysosomal pathway for receptor downregulation. This reliance makes them particularly suitable for probing ANKRD13A??s role in endocytic trafficking within a cancer-relevant context.
ANKRD13A functions as a ubiquitin-binding adaptor that couples ubiquitinated cargo to the ESCRT machinery, directing it into intraluminal vesicles of multivesicular bodies for lysosomal degradation. It directly interacts with ubiquitin, ESCRT-0 components Hrs and STAM, and ESCRT-I subunits Tsg101 and Vps37A. Upon EGF stimulation, ANKRD13A recognizes ubiquitinated EGFR and facilitates its ESCRT-dependent sorting, terminating receptor signaling. Knockout of ANKRD13A consequently impairs EGFR degradation, resulting in sustained phospho-EGFR levels and prolonged activation of downstream pathways that influence cell proliferation, migration, and survival.
In the A-549 lung adenocarcinoma background, ANKRD13A knockout is particularly informative because EGFR signaling is a central oncogenic driver. Aberrant receptor degradation can sustain pro-survival signals even in the presence of tyrosine kinase inhibitors, contributing to therapeutic resistance. This polyclonal knockout model therefore enables detailed interrogation of how ANKRD13A loss reshapes EGFR trafficking dynamics and intersects with the broader signaling networks governing lung cancer cell behavior.
Key experimental applications include cycloheximide chase assays to measure EGFR half-life, co-immunoprecipitation of ubiquitinated receptor complexes, and immunofluorescence co-localization studies of endosomal markers. Surface EGFR retention can be quantified by flow cytometry, while downstream signaling is assessed via phospho-EGFR ELISA or western blotting. The cells also support functional genomics of ubiquitin-dependent sorting, ESCRT pathway dissection, and drug resistance mechanism studies, often combined with viability and migration assays. For additional information, please contact Ascent Research.
