The ASRGL1 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 143B human osteosarcoma cell line, featuring targeted disruption of the ASRGL1 gene. This pooled population contains heterogeneous gene-disrupting edits introduced by CRISPR/Cas9, leading to loss of ASRGL1 expression without clonal isolation. The knockout cells provide a versatile tool for studying ASRGL1 function in a cancer-relevant background.
The host 143B cell line is a tumorigenic, TK-negative osteosarcoma model originating from bone tissue. Widely used in oncology research, 143B cells exhibit aggressive growth and a well-defined genetic profile, making them suitable for metabolic and signaling studies. The bone-derived context offers a physiologically appropriate system for examining cancer cell metabolism, particularly pathways relevant to osteosarcoma.
ASRGL1 encodes a cytosolic L-asparaginase that hydrolyzes L-asparagine to L-aspartate and ammonia, key metabolites in amino acid catabolism. This enzyme is transcriptionally regulated by ATF4 downstream of the PERK-eIF2?? stress-sensing arm, linking nutrient availability to the unfolded protein response (UPR). ASRGL1 activity modulates intracellular asparagine pools, which in turn influence mTORC1 signaling and TCA cycle intermediate levels. Knockout of ASRGL1 ablates this regulatory node, potentially altering GCN2-ATF4 feedback and mTORC1-dependent proliferation. Interplay with ASNS further underscores its role in asparagine homeostasis.
In the 143B osteosarcoma context, ASRGL1 disruption carries particular relevance for studying metabolic vulnerabilities. Osteosarcoma cells rely on amino acid metabolism to sustain growth under nutrient stress, and loss of ASRGL1 may sensitize them to asparagine deprivation or L-asparaginase therapy. This model enables dissection of intrinsic resistance mechanisms to L-asparaginase, a treatment explored beyond leukemia. By eliminating endogenous asparaginase activity, researchers can investigate metabolic reprogramming and stress adaptation, providing insights into therapeutic strategies targeting osteosarcoma metabolism.
Research applications include Western blotting and RT-qPCR for expression analysis, enzyme activity assays to confirm functional KO, and cell proliferation assays under asparagine limitation. Metabolic profiling by LC-MS and L-asparaginase sensitivity assays further characterize metabolic dependencies. These tools facilitate studies on amino acid metabolism, drug resistance, and osteosarcoma biology. For ordering and technical support, contact Ascent Research.
