The AATK Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the AATK gene, encoding the serine/threonine kinase AATK, has been disrupted using CRISPR/Cas9-mediated gene targeting. This product provides a loss-of-function model for studying AATK-dependent signaling pathways in a human cell background, without the need for single-cell cloning. The polyclonal nature ensures that the population retains the phenotypic heterogeneity of the parental HEK293T line, making it suitable for pooled functional genomics and population-level assays that capture a broad spectrum of knockout events.
HEK293T is a derivative of the HEK293 human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen, enabling high-copy episomal replication of plasmids containing the SV40 origin of replication. This feature makes HEK293T an ideal host for transient transfection, high-level recombinant protein expression, and lentiviral or retroviral packaging. In the context of the AATK knockout, HEK293T provides a tractable and extensively characterized model system that supports robust biochemical analyses while avoiding confounding effects from neuronal lineage-specific programs.
AATK (apoptosis-associated tyrosine kinase) is a serine/threonine kinase that functions downstream of the neurotrophin receptor TrkA. Upon nerve growth factor (NGF) stimulation, AATK associates with TrkA and becomes activated, initiating signaling through the JNK mitogen-activated protein kinase pathway. Activated JNK subsequently phosphorylates 14-3-3 proteins and cytoskeletal substrates such as actin and cofilin, leading to cytoskeletal reorganization. In parallel, JNK activation can trigger caspase-3 cleavage and promote apoptosis. Thus, AATK serves as a key node that integrates extracellular neurotrophic signals to coordinate cell morphology and survival decisions.
The HEK293T background, though of renal origin, retains the core components required for AATK signal transduction, enabling the dissection of AATK-dependent mechanisms in a simplified cellular environment. This model is particularly useful for studying how AATK interfaces with TrkA and JNK signaling independent of the full neuronal differentiation machinery. Moreover, the polyclonal knockout strategy mitigates clonal artifacts and allows assessment of gene function at the population level, providing a robust platform for high-throughput screening and quantitative biology approaches.
Typical research applications include immunoblotting to assess AATK protein levels and the phosphorylation status of downstream effectors such as JNK and caspase-3, Annexin V-based apoptosis assays to quantify AATK-dependent cell death, in vitro kinase activity measurements, and co-immunoprecipitation experiments to probe interactions with TrkA or 14-3-3 proteins. Quantitative RT-PCR can be employed to monitor AATK-regulated transcriptional changes. These assays facilitate detailed mechanistic dissection of AATK signaling and enable compound screening for kinase inhibitors. For further information or technical support, please contact Ascent Research.
