The BOLA2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa cell line, designed to achieve functional disruption of the BOLA2 gene. This product provides a loss-of-function model for investigating the role of BOLA2 in iron-sulfur (Fe-S) cluster biogenesis and its broader impacts on cellular metabolism and stress responses. The polyclonal format ensures representation of a heterogeneous knockout pool, suitable for pooled functional studies without requiring single-cell clonal isolation.
HeLa cells were originally established in 1951 from a cervical adenocarcinoma in a 31-year-old African American patient. This widely used epithelial cancer cell line is known for its rapid proliferation, robustness, and extensive characterization in cancer biology, signal transduction, and drug discovery. HeLa cells serve as an appropriate host for studying mitochondrial dysfunction and metabolic reprogramming due to their active oxidative and glycolytic metabolism.
BOLA2 encodes a scaffold protein that functions as a critical component of the iron-sulfur cluster assembly machinery. Mechanistically, BOLA2 forms a heterodimeric complex with glutaredoxin 5 (GLRX5) to mediate the transfer of nascent Fe-S clusters to apoproteins. This process is essential for the maturation of Fe-S cluster-dependent enzymes, including mitochondrial aconitase and Complex I subunits of the respiratory chain. BOLA2 activity is transcriptionally regulated by upstream factors NRF2 and ATF4, which are activated under oxidative stress and iron depletion conditions. In turn, BOLA2 interacts with NFU1, BOLA1, and BOLA3 within a network that also includes FXN, ISCU, NFS1, and ISD11, collectively orchestrating Fe-S cluster delivery to both mitochondrial and cytosolic targets.
Knockout of BOLA2 in HeLa cells leads to impaired Fe-S cluster biogenesis, resulting in defects in mitochondrial respiratory chain function and elevated reactive oxygen species (ROS) levels. This model is highly relevant for studying the intersections between mitochondrial homeostasis and cancer cell metabolism. Given the role of Fe-S clusters in DNA repair and replication, BOLA2 knockout also provides insights into genome stability mechanisms. Moreover, disruptions in Fe-S cluster assembly are implicated in neurodegenerative disorders such as Friedreich’s ataxia, making this model valuable for exploring disease-relevant pathways in an immortalized cell background.
Researchers can utilize these polyclonal knockout cells to investigate BOLA2-dependent processes using a variety of approaches: western blotting to confirm BOLA2 depletion, RT-qPCR for transcriptional analysis, immunofluorescence for mitochondrial morphology assessment, and Seahorse metabolic flux analysis to measure oxidative phosphorylation and glycolysis. Additionally, ROS detection assays, iron staining, and viability assays enable functional characterization of the knockout phenotype. For further inquiries or to discuss customized research applications, please contact Ascent Research.
