DNAJC6 Knockout A2780 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of A2780 cells carrying targeted disruption of the DNAJC6 gene. This loss-of-function model is designed to abolish functional auxilin expression, enabling systematic investigation of its role in clathrin-mediated endocytosis and related cellular processes. The knockout pool is generated by CRISPR/Cas9-mediated genome editing, introducing targeted gene disruption without the need for clonal selection. The polyclonal nature of the pool preserves genetic diversity while ensuring robust target-gene disruption, making it suitable for bulk functional assays.
The A2780 cell line is a well-characterized human ovarian carcinoma model established from an untreated patient. Widely employed in drug sensitivity and resistance studies, this epithelial cancer line retains key signaling pathways relevant to ovarian tumor biology. Its use as host for DNAJC6 knockout provides a clinically pertinent context for evaluating how endocytic trafficking influences cancer cell behavior, including response to chemotherapeutic agents.
DNAJC6 encodes auxilin, a neuronal and non-neuronal J-domain co-chaperone that recruits Hsc70 (HSPA8) to clathrin-coated vesicles. Auxilin binds to clathrin heavy chain (CLTC) and stimulates the ATPase activity of Hsc70, driving the disassembly of the clathrin lattice during vesicle uncoating. This critical step in clathrin-mediated endocytosis is tightly coordinated with dynamin?1 (DNM1) and the adaptor protein complex 2 (AP2), and controls the internalization of numerous cell surface receptors including epidermal growth factor receptor (EGFR) and transferrin receptor (TFRC). Disruption of DNAJC6 therefore impairs clathrin uncoating, stalling endocytic traffic and potentially altering downstream signaling cascades initiated by these receptors.
In A2780 ovarian cancer cells, DNAJC6 knockout is expected to perturb endocytic uptake of oncogenic receptors, thereby modulating proliferative and survival signaling pathways. Aberrant receptor trafficking is increasingly implicated in cancer progression and chemoresistance, and the A2780 model, known for its utility in drug sensitivity profiling, offers a powerful system to dissect these connections. By ablating auxilin function, researchers can directly assess how clathrin-mediated endocytosis contributes to the cellular response to platinum-based chemotherapies and other agents commonly used in ovarian cancer treatment.
This knockout cell population is ideally suited for a range of experimental applications, including quantitative transferrin uptake assays to measure endocytic rate, immunofluorescence staining of clathrin structures, and EGFR internalization kinetics using labeled ligands or pH-sensitive probes. Additionally, western blotting for clathrin and Hsc70 levels, MTS viability assays, Annexin V apoptosis assays, and clonogenic survival assays can be employed to relate endocytic function to drug sensitivity and cell fate. These tools support research into chemoresistance mechanisms, receptor trafficking dynamics, and the broader role of endocytosis in ovarian cancer biology. For further information or to discuss custom projects, please contact Ascent Research.
