The ASCC1 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered for targeted disruption of the ASCC1 gene locus. This loss-of-function model enables investigation of ASCC1-dependent mechanisms without the need for transient silencing approaches. The polyclonal format preserves heterogeneous editing events across the population, providing a robust model for studying ASCC1 function in bulk cellular assays.
These cells are derived from the HEK293T human embryonic kidney epithelial cell line, which constitutively expresses the SV40 large T antigen. This host background supports high transfection efficiency and robust episomal replication, making it a widely adopted system for protein expression, viral production, and functional genomic studies. The adherent epithelial morphology and rapid doubling time facilitate reproducible experimental workflows.
ASCC1 encodes a scaffold subunit of the activating signal cointegrator 1 (ASCC) complex. Upon DNA alkylation damage, the ASCC complex, comprising ASCC1, ASCC2, ASCC3, and TRIP4, is recruited to lesion sites. ASCC1 mediates assembly of the complex and facilitates interaction with RNA polymerase II. The complex subsequently recruits the alkylation repair demethylase ALKBH3, which removes methyl adducts from DNA bases, enabling transcription bypass and promoting cell survival. This pathway is activated upstream by ATM/ATR kinases in response to DNA lesions, and its disruption sensitizes cells to alkylating agents such as methyl methanesulfonate (MMS).
In the HEK293T background, ASCC1 knockout allows direct functional dissection of transcription-coupled repair of alkylated DNA. This model is particularly relevant for studying the cellular response to genotoxic stress in kidney-derived epithelial cells. Given the association of ASCC1 mutations with spinal muscular atrophy with congenital bone fractures, arthrogryposis, and intellectual disability, the knockout cells provide a platform for exploring the molecular pathogenesis of these disorders, despite the non-disease origin of the host line.
Key applications include mechanistic studies of DNA alkylation repair, high-content screening for modulators of alkylation sensitivity, and functional characterization of ASCC1-interacting factors. Researchers can monitor DNA damage accumulation via ??H2AX immunofluorescence, assess repair kinetics with comet assays, and quantify survival after alkylation challenge using colony formation or viability assays. Gene expression changes in downstream targets can be analyzed by RT-qPCR. For further technical details or bulk pricing inquiries, please contact Ascent Research.
