The AGL Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with disrupted AGL gene function. This provides a loss-of-function model for glycogen debranching enzyme studies. The polyclonal format ensures a heterogeneous pool of edited cells without clonal selection, suitable for investigating glycogen metabolism in a tractable cell background.
The parental HEK293T cell line is an adherent human embryonic kidney epithelial line stably expressing SV40 large T antigen, derived from HEK293 cells. It is widely used for recombinant protein expression and viral vector production due to high transfectability. Although not a primary metabolic cell type, HEK293T retains functional glycogen pathways, making it suitable for probing glycogen homeostasis.
AGL encodes the glycogen debranching enzyme, a dual-function enzyme that hydrolyzes ??-1,6-glycosidic bonds and transfers maltotriose residues to acceptor chains during glycogen degradation. It operates in concert with glycogen phosphorylase (PYGL, PYGM, PYGB) and phosphoglucomutase (PGM1) to liberate glucose-1-phosphate and free glucose. AGL activity is tightly controlled by hormonal signals: insulin represses glycogenolysis, while glucagon and epinephrine activate it through the cAMP/PKA cascade. Disruption of AGL prevents branch removal, leading to phosphorylase-limit dextrin accumulation and defective glucose release. This disrupts cellular energy balance and glycogen turnover. Additional pathway components include glucose-6-phosphatase (G6PC) and glycogen synthase (GYS1/GYS2), positioning AGL centrally in glycogen metabolism.
In the HEK293T background, AGL knockout creates a cellular model for glycogen storage disease type III (GSDIII, Cori/Forbes disease), where loss-of-function mutations cause tissue glycogen accumulation. These polyclonal cells exhibit impaired glycogen degradation, recapitulating hallmark features such as aberrant glycogen deposits visible by PAS staining. Although HEK293T are not hepatocytes, they provide a tractable system for studying the molecular pathology of AGL deficiency and for high-throughput screening of small-molecule therapies or pharmacological chaperones. The polyclonal nature better reflects the heterogeneous mutation landscape observed in patients, supporting genotype?Cphenotype correlation analyses.
Research applications encompass modeling GSDIII, dissecting glycogen metabolism regulation, and screening therapeutic candidates. Recommended assays include glycogen content measurement, PAS staining, debranching enzyme activity, Western blotting, RT-qPCR, immunofluorescence for glycogen, and glucose uptake/production measurements. These cells also facilitate interrogation of upstream regulators such as insulin, glucagon, and epinephrine, as well as downstream metabolic impacts. For further information, please contact Ascent Research.
