The ASRGL1 Knockout KYSE-150 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human esophageal squamous cell carcinoma line KYSE-150. This product features targeted disruption of the ASRGL1 gene via CRISPR/Cas9-mediated gene editing, resulting in a heterogeneous pool of cells harboring diverse loss-of-function mutations. The knockout model provides a powerful tool for dissecting the role of ASRGL1 in cancer cell metabolism and tumorigenesis without relying on single-cell clonal isolates.
The parental KYSE-150 cell line is a poorly differentiated esophageal squamous cell carcinoma established from a 49-year-old Japanese female. KYSE-150 cells exhibit aggressive tumorigenic properties and are widely employed as an in vitro and in vivo model for esophageal cancer research. The cell line retains key molecular characteristics of the original tumor, making it a relevant platform for studying oncogenic mechanisms and evaluating therapeutic vulnerabilities.
ASRGL1 encodes an enzyme with dual L-asparaginase and isoaspartyl peptidase activities, catalyzing the hydrolysis of L-asparagine to L-aspartate and ammonia, and cleaving isoaspartyl peptides to facilitate protein repair. The enzyme is activated by amino acid deprivation and L-asparagine levels, functioning downstream of metabolic stress signals. L-aspartate produced by ASRGL1 serves as a substrate for aspartate transaminase and contributes to mTOR pathway regulation, linking ASRGL1 activity to amino acid sensing and anabolic growth. Through these molecular interactions, ASRGL1 plays a central role in asparagine catabolic processes and maintenance of amino acid homeostasis.
In the context of KYSE-150 cells, CRISPR/Cas9-mediated knockout of ASRGL1 disrupts asparagine hydrolysis and isoaspartyl peptide cleavage, impairing aspartate production and protein repair. This metabolic perturbation is predicted to alter mTOR signaling and reduce cellular proliferation, underscoring the dependency of esophageal cancer cells on ASRGL1-driven metabolism. The polyclonal knockout population thus provides a physiologically relevant model to study amino acid dependencies and metabolic adaptations in esophageal squamous cell carcinoma.
These polyclonal knockout cells are optimized for functional studies of ASRGL1 in tumorigenesis and for drug target validation focused on amino acid dependency. Researchers can employ western blotting and RT-qPCR to confirm ASRGL1 disruption and assess compensatory pathways. Proliferation and apoptosis can be monitored via MTT and apoptosis assays, while colony formation assays evaluate long-term survival. Amino acid quantification enables direct measurement of altered L-asparagine and L-aspartate levels, offering insights into metabolic rewiring. For further information, please contact Ascent Research.
