CRISPR/Cas9-mediated disruption of the ASRGL1 gene in T-47D cells generates a polyclonal knockout cell population that serves as a loss-of-function model for studying L-asparaginase activity in hormone-responsive breast cancer. This product consists of a heterogeneous pool of edited cells, allowing researchers to assess the overall impact of ASRGL1 inactivation without clonal selection artifacts, and is supplied as a ready-to-use polyclonal knockout cell stock for downstream functional assays.
The T-47D host cell line is an estrogen receptor (ER)-positive, progesterone receptor (PR)-positive, and HER2-negative breast ductal carcinoma epithelial model originally derived from a pleural effusion. These cells represent the luminal A molecular subtype and are widely employed to investigate hormone-dependent signaling, endocrine therapy responses, and the metabolic adaptations of breast cancer cells.
ASRGL1 encodes an L-asparaginase that hydrolyzes L-asparagine to L-aspartate and ammonia, playing a critical role in amino acid homeostasis and protein turnover. Its activity is regulated by estrogen receptor signaling, the stress-responsive transcription factor ATF4, and nutrient deprivation. Downstream, ASRGL1-mediated asparagine depletion inhibits mTORC1 signaling, promotes caspase-3 activation, and induces G1/S cell cycle arrest. ASRGL1 forms a functional homodimer and directly interacts with its asparagine substrate, linking amino acid metabolism to cell growth and survival pathways.
In T-47D cells, knockout of ASRGL1 abolishes L-asparaginase activity, leading to elevated intracellular asparagine levels that enhance mTORC1 signaling and suppress apoptosis. This metabolic reprogramming may promote proliferation and contribute to endocrine therapy resistance, highlighting ASRGL1??s tumor-suppressive function in hormone-responsive breast cancer. The polyclonal knockout model enables the study of how loss of asparagine-degrading capacity influences estrogen receptor-driven growth and tamoxifen sensitivity.
This knockout cell product is suitable for a broad range of research applications, including profiling amino acid metabolism and its interplay with mTOR signaling in breast cancer, investigating mechanisms of endocrine resistance such as tamoxifen resistance, identifying metabolic vulnerabilities through synthetic lethality screens, and evaluating apoptosis pathway dynamics. Compatible assays include Western blotting for mTORC1 targets, asparaginase activity measurements, cell proliferation and apoptosis assays, phospho-S6 kinase analysis, LC-MS-based metabolomics, and drug sensitivity profiling with asparaginase. For ordering, technical support, or inquiries, please contact Ascent Research.
