The ASRGL1 Knockout PaTu 8988t Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the human ASRGL1 gene. This heterogeneous pool of edited cells provides a robust loss-of-function model, circumventing clonal variability and enabling population-level analyses of gene function. The polyclonal format is particularly suited for pooled screens and studies where biological noise must be minimized while preserving diverse editing outcomes.
The host cell line, PaTu 8988t, is an established model of human pancreatic ductal adenocarcinoma (PDAC) derived from a liver metastasis. These cells carry a KRAS G12V oncogenic mutation, a hallmark of aggressive PDAC with constitutive activation of downstream proliferative and survival pathways. PaTu 8988t cells retain key features of pancreatic exocrine secretion and metastatic capacity, making them a physiologically relevant system for probing metabolic adaptations in advanced pancreatic cancer.
ASRGL1 encodes a bifunctional enzyme with asparaginase and isoaspartyl dipeptidase activities, catalyzing the hydrolysis of L-asparagine to aspartic acid and ammonia, and repairing isoaspartyl damage in proteins. The enzyme functions downstream of amino acid deprivation signals and is transcriptionally regulated by ATF4 during the integrated stress response. Its product, aspartate, and the depletion of asparagine intersect with mTORC1 signaling, which integrates glutamine and aspartate levels to govern protein synthesis and cell growth. The network includes asparagine synthetase (ASNS), glutamine, and mTORC1 as representative components, with ASRGL1 acting as a critical node linking protein repair to metabolic control.
In the context of KRAS-mutant PDAC, ASRGL1 disruption is expected to perturb asparagine homeostasis and protein integrity maintenance. Pancreatic cancer cells often display heightened dependency on asparagine, and the loss of ASRGL1 may exacerbate metabolic stress, potentially reducing proliferation and increasing sensitivity to asparagine-depleting agents like asparaginase. Furthermore, impaired isoaspartyl repair could lead to accumulation of damaged proteins, triggering proteotoxic stress and altering apoptotic thresholds. This knockout model thus offers a platform to dissect how oncogenic KRAS cooperates with amino acid metabolism and protein quality control pathways to sustain tumor growth and metastasis.
Key applications include investigating the role of asparagine metabolism in PDAC, evaluating resistance mechanisms to asparaginase therapy, and exploring metabolic crosstalk between ASRGL1, ASNS, and mTOR signaling. Typical assays encompass immunoblotting for ASRGL1 and downstream targets, quantification of asparaginase activity in lysates, cell viability (MTS/MTT) and apoptosis assays under asparagine deprivation, LC-MS-based amino acid profiling, colony formation assays, and drug sensitivity testing with asparaginase. RT-qPCR can measure changes in ATF4, ASNS, and mTOR pathway genes. For additional product information or technical support, please contact Ascent Research.
