The ASRGL1 Knockout Ca Ski Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ASRGL1 gene in the Ca Ski human cell line. This product provides a heterogeneous pool of gene-disrupted cells, enabling researchers to study loss-of-function effects without clonal biases. CRISPR/Cas9-mediated disruption of ASRGL1 abrogates expression of the bifunctional L-asparaginase/isoaspartyl peptidase, making it a valuable tool for investigating asparagine metabolism, protein repair pathways, and their roles in cervical cancer biology.
The Ca Ski host cell line, derived from a cervical epidermoid carcinoma metastasis to the small intestine mesentery, is a widely employed model for HPV-16 positive cervical cancer. Ca Ski cells harbor an integrated HPV-16 genome and exhibit adherent epithelial morphology. Their sustained oncogenic signaling and well-characterized molecular landscape establish them as a relevant system to explore metabolic vulnerabilities and therapeutic responses, particularly in the context of HPV-driven malignancies.
ASRGL1 encodes a bifunctional enzyme that hydrolyzes L-asparagine to L-aspartate and ammonia, and also repairs damaged proteins through isoaspartate cleavage. This enzymatic activity positions ASRGL1 at a critical node in amino acid homeostasis. Upstream, ASRGL1 expression is regulated by the integrated stress response effector ATF4, the MYC transcriptional program, and nutrient deprivation. Downstream, ASRGL1-mediated asparagine depletion can inhibit mTORC1 signaling, reducing phosphorylation of S6K and 4E-BP1, and suppress protein synthesis. ASRGL1 functionally interacts with ASNS (asparagine synthetase) and participates in metabolic enzyme complexes that balance asparagine levels, thereby influencing cell proliferation and survival under metabolic stress.
In Ca Ski cervical carcinoma cells, knockout of ASRGL1 disrupts the intricate control of intracellular asparagine, potentially mimicking asparaginase treatment and sensitizing cells to metabolic stress. This model allows dissection of how HPV-16 oncoproteins may crosstalk with amino acid sensing pathways via ATF4 and mTORC1. The polyclonal nature captures diverse mutational outcomes, offering a robust platform to assess phenotypic heterogeneity in drug sensitivity, particularly to L-asparaginase-based therapies, and to identify resistance mechanisms that could arise in acute lymphoblastic leukemia and other L-asparaginase-resistant malignancies.
Researchers can apply these polyclonal knockout cells in a broad range of experimental contexts, including western blotting to verify ASRGL1 loss and assess mTORC1 effectors (e.g., phospho-S6), RT-qPCR for transcript-level confirmation, and enzymatic assays to quantify asparaginase activity. Functional studies may incorporate MTT proliferation assays, Annexin V apoptosis assays, cell cycle analysis, and colony formation assays under varying asparagine concentrations or drug treatments. Metabolomic profiling and asparagine/aspartate quantification further enable detailed metabolic flux analyses. This product is ideally suited for studies on chemoresistance, biomarker discovery, and the development of L-asparaginase combination therapies. For additional information, validation data, or custom knockout services, please contact Ascent Research.
