The ATM Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted disruption of the ATM gene. This polyclonal pool, derived from HEK293T cells, provides a loss-of-function model that avoids single-cell cloning, preserving the inherent genetic heterogeneity of a polyclonal population. CRISPR/Cas9-mediated gene editing yields a functional knockout, abolishing ATM kinase activity without introducing clonal artifacts, making it suitable for studies where population-level responses are representative of the bulk cellular context.
HEK293T cells are a human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen, enabling episomal plasmid amplification and robust protein production. They exhibit adherent epithelial morphology and are extensively employed as a model system for renal epithelial biology, signal transduction, and heterologous gene expression. Their high transfectability and genetic tractability facilitate efficient generation of gene-edited derivatives, establishing a versatile platform for dissecting molecular pathways in a diploid human cellular environment.
ATM encodes a serine/threonine protein kinase that serves as a master regulator of the DNA damage response, primarily activated by DNA double-strand breaks (DSBs) through the MRN complex (MRE11?CRAD50?CNBS1) and chromatin alterations. Upon activation, ATM phosphorylates a cascade of downstream targets, including p53, CHK2, BRCA1, H2AX (yielding ??-H2AX), NBS1, SMC1, KAP1, and MDM2, thereby coordinating cell cycle checkpoints, DNA repair via homologous recombination and non-homologous end joining, and apoptosis. It also engages in complex interactions with ATR, DNA-PKcs, Tip60, PP2A phosphatase, and BRCA1, integrating signals from oxidative stress and auto-phosphorylation to maintain genomic integrity.
Within the HEK293T background, disruption of ATM eliminates the principal kinase activity that transduces DSB signals, severely compromising G1/S and G2/M cell cycle arrest, diminishing p53-dependent transcriptional responses, and reducing formation of ??-H2AX and 53BP1 nuclear foci. As a result, these polyclonal knockout cells display marked sensitivity to ionizing radiation and DNA-damaging chemotherapeutics, recapitulating hallmark features of ATM deficiency. This model enables detailed examination of ATM-dependent and -independent DNA repair mechanisms in a human epithelial context, facilitating research into synthetic lethality and cancer vulnerabilities.
Researchers can apply this product in a wide range of experimental workflows, including Western blot analysis of total and phosphorylated ATM, CHK2, and ??-H2AX; immunofluorescence microscopy to quantify damage-induced foci; comet assays for direct assessment of DNA strand breaks; flow cytometric profiling of cell cycle distribution and apoptosis; and clonogenic survival assays following genotoxic stress. It is also suited for drug screening campaigns targeting DNA repair pathways, genome stability assays using fluorescent reporters, and RT-qPCR quantification of downstream transcriptional changes such as CDKN1A (p21) and BAX. For additional information or to discuss your specific project needs, please contact Ascent Research.
