The ATE1 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-edited population of HT29 human colorectal adenocarcinoma epithelial cells with targeted disruption of the ATE1 gene. This polyclonal knockout pool comprises a heterogeneous collection of edited cells, minimizing clonal selection artifacts and enabling robust loss-of-function analysis in a genetically diverse background. The model allows researchers to study the functional consequences of abrogated ATE1 activity in a transformed intestinal epithelial milieu.
HT29 cells, derived from a primary adenocarcinoma of the colon, are a well-established model for investigating intestinal epithelial barrier function and colorectal cancer biology. These cells harbor mutations in key tumor suppressors such as APC and TP53, form polarized monolayers, and are widely employed in studies of oncogenic signaling, drug transport, and metastasis. Their colonic origin makes them particularly relevant for dissecting pathways that contribute to colorectal tumorigenesis and therapeutic resistance.
ATE1 catalyzes the post?translational arginylation of N?terminal aspartate, glutamate, or oxidized cysteine residues, marking substrate proteins for recognition by UBR1 and UBR2 E3 ubiquitin ligases and subsequent degradation by the 26S proteasome. Key arginylation targets include beta-actin, calreticulin, and alpha-synuclein, linking ATE1 to cytoskeletal dynamics, ER protein quality control, and proteostasis. The enzyme is activated by upstream stress signals transduced through ATF4 and HIF1A transcription factors and calcium cascades, and it functionally cooperates with arginyl?tRNA synthetase (RARS), molecular chaperones, and proteasome components to fine?tune protein turnover under changing cellular conditions.
Disruption of ATE1 in the HT29 colorectal cancer model ablates N?end rule-mediated degradation, leading to aberrant stabilization of arginylation substrates and altered stress signaling networks. This perturbation allows researchers to deconvolve the contribution of arginylation to processes such as proliferation, apoptosis, epithelial?mesenchymal transition, and drug sensitivity. The knockout model thus serves as a powerful tool for understanding how dysregulation of the N?end rule pathway promotes colon cancer progression and influences response to chemotherapeutics.
The polyclonal ATE1 knockout pool is suitable for a broad range of experimental applications, including western blotting to monitor substrate accumulation, arginylation activity assays, RT?qPCR for transcriptional analysis, cell viability and apoptosis profiling under drug treatment, and proteasome activity measurements. It is also well suited for pooled drug screens and functional migration/invasion assays, providing a versatile system to interrogate N?end rule biology in colorectal cancer. For additional product information or technical inquiries, please contact Ascent Research.
