The BOLA1 Knockout A-549 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A-549 lung adenocarcinoma epithelial cell line. This product comprises a pool of cells harboring targeted disruptions in the BOLA1 gene, generated via CRISPR/Cas9-mediated gene editing without clonal isolation. The polyclonal format provides a heterogeneous mixture of knockout alleles, enabling loss-of-function studies of BOLA1 in a non-small cell lung cancer (NSCLC) model.
The parental A-549 cell line is a widely employed model of NSCLC, established from a 58-year-old male patient. These adherent epithelial cells retain wild-type p53 functionality and are extensively characterized for oncogenic signaling, drug response, and tumor biology research. A-549 cells serve as a robust platform for investigating the molecular mechanisms underlying lung adenocarcinoma, particularly in the context of mitochondrial metabolism, oxidative stress, and apoptosis regulation.
BOLA1 encodes a mitochondrial protein critical for iron-sulfur (Fe-S) cluster biogenesis, where it interacts directly with GLRX5 to mediate Fe-S transfer to client proteins, including respiratory chain complexes and aconitase. The BOLA1/GLRX5 axis operates downstream of the ISCU?CNFS1?CFXN core assembly machinery and cooperates with other late-acting factors such as NFU1, BOLA3, ISCA1, and ISCA2. In A-549 cells, BOLA1 expression is regulated by oxidative stress-responsive transcription factor NRF2, the tumor suppressor p53, and oncogenic KRAS signaling. Genomic disruption of BOLA1 impairs mitochondrial oxidative phosphorylation, elevates intracellular reactive oxygen species (ROS), and activates pro-apoptotic cascades involving upregulation of BAX, downregulation of BCL-2, and cleavage of caspase-3. Additionally, downstream targets include the cell cycle inhibitor p21, linking Fe-S metabolism to proliferation control.
In the A-549 background, BOLA1 knockout provides a unique tool to dissect the interplay between mitochondrial iron metabolism and hallmark cancer phenotypes. Loss of BOLA1 is expected to sensitize NSCLC cells to mitochondrial stress and lower the threshold for apoptosis induction, particularly under conditions of elevated oxidative load or chemotherapeutic challenge. This model enables precise investigation of how Fe-S cluster trafficking influences tumor cell fitness, metabolic plasticity, and treatment resistance, offering insights into potential therapeutic vulnerabilities in lung adenocarcinoma.
Key applications include mechanistic studies of BOLA1 in lung adenocarcinoma pathogenesis, evaluation of iron-sulfur cluster metabolism in cancer, and assessment of mitochondrial dysfunction and ROS-mediated apoptosis. The polyclonal knockout pool is amenable to cell viability assays (MTT), colony formation, ROS detection (H2DCFDA), mitochondrial membrane potential measurement (JC-1), aconitase activity quantification, and Seahorse respirometry. Further analyses by Western blotting may assess expression changes of BOLA1, GLRX5, cleaved caspase-3, and BCL-2 family proteins, while flow cytometry facilitates apoptosis (Annexin V/PI) and cell cycle analyses. For detailed product inquiries or technical support, please contact Ascent Research.
