The ASRGL1 Knockout HT29 Polyclonal Cells comprise a genetically heterogeneous population of HT29 colorectal adenocarcinoma cells engineered via CRISPR/Cas9-mediated gene disruption to ablate ASRGL1 expression. This polyclonal knockout model retains the inherent diversity of a non-clonal edited pool, enabling robust assessment of ASRGL1 loss-of-function within a physiologically relevant epithelial tumor context without the biases associated with single-cell clonal selection.
The HT29 parental cell line originates from a primary colorectal adenocarcinoma of a 44-year-old female and is widely employed as a model system for intestinal epithelial biology and colorectal cancer research. These adherent epithelial cells retain characteristics of enterocytic differentiation, including polarization and brush-border enzyme expression under specific conditions, making them a valuable platform for dissecting metabolic dependencies and signaling aberrations in colon carcinoma.
ASRGL1 encodes an L-asparaginase and isoaspartyl dipeptidase that hydrolyzes L-asparagine to L-aspartate and ammonia, serving as a critical regulator of intracellular asparagine homeostasis. In response to amino acid deprivation, the transcription factor ATF4 upregulates ASRGL1 to sustain asparagine supply for nucleotide biosynthesis and mTORC1 signaling. Downstream, ASRGL1 activity supports mTORC1 complex integrity, involving mTOR, Raptor, and S6K1, and promotes protein synthesis while suppressing apoptosis. The enzyme also functionally interacts with ASNS and glutaminase, enzymes that modulate glutamine- and asparagine-dependent anabolic pathways, positioning ASRGL1 at a metabolic crossroad linking amino acid metabolism to cell growth and survival.
In the HT29 colorectal adenocarcinoma model, disruption of ASRGL1 creates a distinct metabolic vulnerability by depleting intracellular aspartate pools and blunting mTORC1 activity. This loss-of-function state impairs proliferation and triggers apoptotic programs, closely mimicking the anti-neoplastic effects of L-asparaginase therapy observed in hematological malignancies. Consequently, these knockout cells provide a human solid-tumor platform to dissect how asparagine metabolism interfaces with oncogenic signaling networks in colorectal cancer, offering insight into tissue-specific metabolic adaptations that distinguish solid tumors from leukemias.
The ASRGL1 Knockout HT29 Polyclonal Cells are optimally suited for a spectrum of mechanistic and translational studies, including western blotting for ASRGL1 and phospho-S6K1, RT-qPCR, MTT and Annexin V-based proliferation/apoptosis assays, LC-MS metabolomic profiling of asparagine and aspartate, colony formation analyses, and xenograft tumor growth experiments. Researchers can employ this model to investigate ASRGL1??s oncogenic role in colorectal cancer, screen for L-asparaginase sensitizers, or dissect mTOR signaling dynamics. For further information or technical support, please contact Ascent Research.
