The BRAT1 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited knockout cell line designed for precise investigation of DNA damage response and ATM signaling. By disrupting the BRAT1 gene, this model creates a permanent loss-of-function system, allowing researchers to dissect BRAT1-dependent mechanisms in genome stability without the confounding effects of transient gene silencing.
The host cell line, HEK293T, is a human embryonic kidney epithelial line widely adopted for its robust growth and high transfection efficiency. Derived from HEK293, it stably expresses the SV40 large T-antigen, facilitating episomal plasmid replication, and harbors adenovirus type 5 sequences, which together support enhanced protein production and viral packaging.
BRAT1 (BRCA1-associated ATM activator 1) is essential for ATM kinase activation following DNA double-strand breaks or ionizing radiation. It interacts with BRCA1 and ATM, promoting phosphorylation of downstream targets such as H2AX (??H2AX at Ser139), CHK2, and p53, thereby enforcing G2/M cell cycle checkpoint and homologous recombination repair. BRAT1 also coordinates with ATR and the MRE11-RAD50-NBS1 (MRN) sensor complex to transduce DNA damage signals. These post-translational modifications orchestrate cell cycle arrest at G2/M and facilitate DNA repair, underscoring BRAT1’s role in maintaining genomic fidelity.
In the HEK293T context, BRAT1 ablation is expected to impair ATM-dependent phosphorylation cascades, leading to defective DNA repair and compromised cell cycle checkpoints. Because HEK293T cells are aneuploid and possess oncogenic stress from adenoviral transformation, this knockout model provides a distinct platform to investigate synthetic lethal interactions and genotoxic drug responses. It is also relevant for modeling neurodevelopmental pathologies linked to BRAT1 mutations, such as lethal neonatal rigidity and multifocal seizure syndrome (RMFSL).
Representative assays include Western blotting for phosphorylated ATM and its substrates, ??H2AX immunofluorescence to visualize DNA damage foci, flow-cytometric cell cycle analysis, and comet assay for DNA break quantification. Researchers can also perform co-immunoprecipitation to probe ATM-BRAT1 interactions and RT-qPCR to assess DNA repair gene expression, enabling comprehensive pathway dissection. This cell line is a rigorous tool for cancer cell biology, drug sensitivity profiling, and disease modeling. For further details or to request a quote, please contact Ascent Research.
