The BRAT1 Knockout HeLa Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa cervical adenocarcinoma cell line, with targeted disruption of the BRAT1 gene. This heterogeneous pool preserves genetic diversity and enables robust functional analyses of BRAT1-dependent DNA damage signaling, cell cycle regulation, and apoptosis without clonal artifacts.
The parental HeLa line, an HPV18-positive aneuploid epithelial cell line from Henrietta Lacks, is a standard model in cancer biology due to its high proliferative index and genetic tractability. HPV E6-mediated p53 degradation partially attenuates canonical p53 responses, yet ATM-dependent signaling remains functional, allowing dissection of BRAT1-mediated ATM regulation in a cervical cancer-relevant context.
BRAT1 enhances ATM kinase activity in response to DNA double-strand breaks by facilitating ATM autophosphorylation and interacting with BRCA1, BARD1, and DNA-PKcs. It promotes phosphorylation of downstream targets: p53 at Ser15, H2AX at Ser139 (??-H2AX), and Chk2 at Thr68, which collectively drive cell cycle arrest, DNA repair, and intrinsic apoptosis. BRAT1 also contributes to mitochondrial homeostasis, linking genotoxic stress to metabolic checkpoints. Through the MRN complex and ATR, BRAT1 integrates damage signals to balance survival and death decisions.
BRAT1 disruption in HeLa cells impairs ATM activation and downstream phosphorylation, sensitizing them to DNA-damaging agents like etoposide and ionizing radiation. This model is relevant for investigating lethal neonatal rigidity and multifocal seizure syndrome (RMFSL), as it reveals how BRAT1 loss affects cell cycle and viability pathways. Additionally, the HPV-positive background clarifies how viral transformation alters dependency on BRAT1-mediated signaling, aiding cancer susceptibility and targeted therapy studies.
These polyclonal knockout cells are suitable for western blotting, RT-qPCR, ??-H2AX immunofluorescence, comet assays, flow cytometry for cell cycle, and annexin V apoptosis assays. Colony formation and drug sensitivity assays with etoposide or IR further characterize the role of BRAT1 in chemoresistance and radiosensitivity. The model also supports mitochondrial and neurodevelopmental disease research when combined with neuronal models. For more information, contact Ascent Research.
