This product is a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ASRGL1 gene in HCT 116 cells. The knockout pool provides a mixed genetic background with disruptions in ASRGL1, suitable for functional studies where clonal variation is less critical. The ASRGL1 locus has been disrupted using CRISPR/Cas9 technology, resulting in a loss-of-function model for investigating the biological roles of this enzyme.
The host cell line, HCT 116, is a widely used human colorectal carcinoma model derived from colon epithelial cells. It harbors a KRAS G13D mutation and exhibits microsatellite instability-high (MSI-H) while maintaining wild-type TP53 status. These characteristics make it a well-characterized system for studying colorectal cancer signaling, drug responses, and tumor suppressor functions.
ASRGL1 encodes an enzyme with L-asparaginase and beta-aspartyl peptidase activities, catalyzing the hydrolysis of L-asparagine to aspartate and ammonia. By depleting intracellular asparagine, ASRGL1 triggers amino acid deprivation responses, including GCN2-mediated phosphorylation of eIF2?? and subsequent activation of the ATF4 transcriptional program. This signaling cascade can inhibit mTORC1 activity, as asparagine is required for mTORC1 activation. The enzyme functionally opposes ASNS, which synthesizes asparagine, thus playing a critical role in asparagine homeostasis. Downstream, ATF4 induces genes involved in stress adaptation, but persistent asparagine depletion can lead to apoptosis in asparagine auxotrophic cells.
In the colorectal cancer context of HCT 116 cells, ASRGL1 may act as a tumor suppressor by limiting asparagine availability and restraining mTORC1-driven growth. The polyclonal knockout model disrupts this regulation, enabling investigation of how asparagine metabolism influences colorectal carcinoma proliferation, survival, and sensitivity to metabolic stress. This model is particularly relevant given the MSI-H background and KRAS mutation, which may intersect with amino acid sensing pathways.
Researchers can employ these knockout cells to study ASRGL1??s role in tumor suppression, asparagine metabolism, and the integrated stress response. Representative applications include Western blotting for ASRGL1 and phospho-eIF2??/ATF4, RT-qPCR for transcript levels, intracellular asparagine quantification, cell proliferation (MTS/MTT) and apoptosis (Annexin V/PI) assays, phospho-S6K1 Western blotting to assess mTORC1 activity, and L-asparaginase sensitivity assays (IC50). These tools support drug discovery and mechanistic studies in colorectal cancer. For further information, please contact Ascent Research.
