The ASB6 Knockout A-549 Polyclonal Cells product comprises a population of CRISPR/Cas9-edited A-549 human lung adenocarcinoma epithelial cells with targeted disruption of the ASB6 gene. As a polyclonal knockout pool, the population retains a heterogeneous array of edited alleles, providing a robust loss-of-function model suitable for functional studies without the need for single-cell cloning. This format enables assessment of gene function at the population level, minimizing clonal bias and reflecting averaged effects of ASB6 loss across the bulk culture.
The A-549 host cell line is derived from the lung adenocarcinoma tissue of a 58-year-old Caucasian male and is widely employed as a model of alveolar type II epithelial cells. These cells maintain key features of lung adenocarcinoma, including their epithelial morphology, proliferative capacity, and responsiveness to growth factors and cytokines. They are extensively used in cancer biology research to study oncogenic signaling, drug metabolism, and tumor cell-intrinsic pathways relevant to lung cancer progression.
ASB6 encodes a substrate recognition component of a Cullin 5-RING E3 ubiquitin ligase complex. Within this complex, ASB6 interacts with Elongin B/C, Cullin 5 (CUL5), and Rbx2 to recruit insulin receptor substrate 4 (IRS4) for ubiquitination and subsequent degradation by the 26S proteasome. By promoting IRS4 turnover, ASB6 acts as a negative regulator of insulin signaling, attenuating the PI3K/AKT cascade downstream of the insulin receptor. ASB6 expression itself is induced by cytokines including interleukin-6 (IL-6) and interferon-gamma (IFN-??), linking JAK-STAT-mediated transcriptional activation to control of insulin sensitivity through ubiquitin-proteasomal degradation.
In the A-549 lung adenocarcinoma context, ASB6 knockout is anticipated to stabilize IRS4 protein levels, thereby potentiating insulin-stimulated PI3K/AKT signaling. This offers a defined system to examine the intersection of ubiquitin-mediated protein turnover and oncogenic metabolic reprogramming. Since A-549 cells express functional insulin receptors and components of the JAK-STAT pathway, this model permits dissection of how cytokine-driven E3 ligase expression influences cancer cell proliferation, survival, and glucose metabolism.
Researchers can employ ASB6 Knockout A-549 Polyclonal Cells to investigate the role of ASB6 in lung adenocarcinoma progression through standard cancer cell biology assays including cell proliferation, migration, and invasion studies. The product is also suited for mechanistic studies of ubiquitin ligase-dependent regulation of IRS4 stability using Western blotting, ubiquitination assays, and co-immunoprecipitation. Downstream signaling events can be profiled via phospho-AKT analysis, while metabolic impacts are assessed through glucose uptake assays, supported by RNA-seq transcriptomics and flow cytometric cell cycle analysis. By combining these approaches, users can validate ASB6 as a therapeutic target in cancer metabolism or metabolic disorders. For technical support and additional information, please contact Ascent Research.
