The ASCC2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the ASCC2 gene in a human embryonic kidney background. This polyclonal product contains a heterogeneous mix of edited cells with targeted disruption of ASCC2, providing a versatile model for examining ASCC2-dependent mechanisms in a cell type renowned for high transfection efficiency and robust protein production.
The host cell line, HEK293T, is a human embryonic kidney epithelial derivative of HEK293 that stably expresses the SV40 large T antigen, enabling episomal replication of plasmids containing the SV40 origin. These cells are extensively employed for transient gene expression, viral packaging, and recombinant protein production due to their high transfectability and robust growth. The constitutive expression of large T antigen enhances their capacity for DNA replication and supports sustained cell division, making HEK293T an optimal host for loss-of-function investigations.
ASCC2 (Activating Signal Cointegrator 1 Complex Subunit 2) is a core component of the ASC-1 transcriptional coactivator complex and a key factor in ribosome-associated quality control (RQC). It facilitates ligand-dependent transcriptional activation by nuclear receptors, such as retinoic acid receptor (RAR) and thyroid hormone receptor (TR), promoting expression of target genes like RAR??2. In RQC, ASCC2 recognizes ubiquitinated nascent chains on stalled ribosomes and recruits the ASCC3 helicase to resolve the complex, directing aberrant polypeptides for proteasomal degradation. ASCC2 thus integrates signals from ligand-activated receptors, DNA damage stimuli, and translational stalling to coordinate gene regulation and protein homeostasis.
Disruption of ASCC2 in HEK293T cells impairs both transcriptional coactivation and RQC. Loss of coactivator function reduces nuclear receptor target gene expression, attenuating hormone-dependent signaling. Concurrently, defective RQC leads to accumulation of misfolded nascent proteins, increasing proteotoxic stress and potentially triggering DNA damage responses and apoptosis. The high translational activity and active ubiquitin-proteasome system of HEK293T cells render these phenotypes particularly pronounced, making this polyclonal knockout model well-suited for investigating the molecular basis of Barrett esophagus and esophageal adenocarcinoma.
Research applications include luciferase reporter assays to assess nuclear receptor transactivation, RT-qPCR for target gene profiling, and western blotting for ASCC2 and interacting proteins. RQC function can be interrogated via ubiquitin affinity purification and proteasome activity assays, while DNA damage signaling is monitored by ??-H2AX foci formation and flow cytometric analysis of cell cycle and apoptosis. These assays facilitate detailed studies in nuclear receptor biology, protein quality control, and disease modeling. For more information or to inquire about this product, please contact Ascent Research.
