The BOLA1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the BOLA1 gene in the human HT29 colorectal adenocarcinoma epithelial cell line. This polyclonal pool provides a loss-of-function model for studying BOLA1-dependent processes without clonal selection artifacts, enabling robust population-level analyses of mitochondrial iron-sulfur cluster biogenesis and redox homeostasis. The product is designed for researchers investigating the interplay between mitochondrial quality control and tumor cell metabolism, offering a ready-to-use tool for functional genomics and drug response studies.
HT29 cells are derived from a human colorectal adenocarcinoma and carry characteristic oncogenic mutations, including BRAF V600E and p53 alterations, which drive constitutive MAPK pathway signaling and impair cell cycle regulation. As an intestinal epithelial model, HT29 cells recapitulate key features of colorectal cancer, such as aberrant proliferation, metabolic reprogramming, and resistance to apoptosis. Their epithelial origin and tumorigenic background make them particularly suitable for dissecting mitochondrial contributions to cancer cell survival under nutrient stress and therapeutic challenges.
BOLA1 encodes a mitochondrial protein that facilitates iron-sulfur (Fe-S) cluster assembly, a process essential for the maturation of Fe-S cluster-containing enzymes such as aconitase (ACO2), respiratory chain complex I subunit NDUFS1, and succinate dehydrogenase subunit B (SDHB). BOLA1 functions within a multiprotein pathway that includes NFS1, ISCU, frataxin (FXN), GLRX5, BOLA3, NFU1, and IND1, and it forms direct complexes with GLRX5, ISCU, and BOLA3. Upstream, BOLA1 expression is transcriptionally regulated by PPARGC1A, NRF1, TFAM, and HIF1A, linking its activity to mitochondrial biogenesis and oxygen sensing. Disruption of BOLA1 impairs Fe-S cluster delivery to client proteins, leading to respiratory chain deficiency and elevated reactive oxygen species (ROS) production, which can trigger apoptosis under stress conditions.
In the context of HT29 cells, BOLA1 knockout provides a powerful system to explore how mitochondrial Fe-S cluster defects influence redox balance, metabolic flexibility, and sensitivity to chemotherapeutic agents. The BRAF V600E-driven oncogenic background of HT29 cells creates a high demand for mitochondrial ATP production and antioxidant defenses; loss of BOLA1 may therefore selectively compromise these pathways, offering a model to study synthetic lethality or druggable vulnerabilities in colorectal cancer. This knockout model also enables investigation of how mitochondrial dysfunction intersects with p53-mediated apoptosis and autophagy in tumor cells.
Research applications include mitochondrial dysfunction studies using Seahorse respirometry and aconitase activity assays, oxidative stress profiling via ROS measurement, and apoptosis assays under conditions of metabolic challenge. RNA-seq and RT-qPCR can be employed to map transcriptional responses to Fe-S cluster deficiency, while western blotting validates downstream targets such as ACO2 and SDHB. The cells are suitable for drug sensitivity screening to identify compounds that exacerbate mitochondrial stress in colorectal cancer models. For additional details, please contact Ascent Research.
