Description

The PGM1 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited knockout cell line designed for targeted disruption of the PGM1 gene in a widely used human embryonic kidney epithelial background. This product provides a defined loss-of-function model in which phosphoglucomutase 1 (PGM1) expression is eliminated through CRISPR/Cas9-mediated gene disruption, enabling systematic investigation of glycogen metabolism, glycolysis, and associated congenital disorders. Researchers can employ this cell line to interrogate PGM1-dependent metabolic pathways, assess substrate flux through central carbon metabolism, and evaluate therapeutic strategies for PGM1 deficiency. The knockout cell line is supplied as a ready-to-use, live cell product suitable for standard cell culture and downstream functional assays.

The host cell line, HEK293T, is a derivative of the HEK293 line that stably expresses the SV40 large T-antigen, permitting episomal replication of plasmids containing the SV40 origin of replication. This feature makes HEK293T exceptionally amenable to transient transfection, lentiviral packaging, and gene editing applications, facilitating complementation studies, rescue experiments, and overexpression of PGM1 variants. The embryonic kidney epithelial origin offers a relevant physiological context for examining glycogen homeostasis and glucose metabolism outside of hepatic or muscle lineages, while the high transfectability ensures efficient delivery of constructs for pathway modulation and metabolic flux analysis.

PGM1 encodes a key enzyme that catalyzes the reversible interconversion of glucose-1-phosphate and glucose-6-phosphate, a critical node linking glycogenolysis and glycogenesis to glycolysis, gluconeogenesis, and the pentose phosphate pathway. PGM1 activity is regulated allosterically by glucose-6-phosphate and is dependent on the cofactor Mg2?. Functionally, PGM1 acts downstream of glycogen phosphorylase (PYGL) and upstream of hexokinase (HK) and glucose-6-phosphatase (G6PC), positioning it at a central metabolic intersection. Disruption of PGM1 ablates the capacity to mobilize glycogen stores, impairing glucose-6-phosphate production and altering downstream pathways that rely on this metabolite, including UDP-glucose synthesis via UDP-glucose pyrophosphorylase (UGP2). This knockout model thus enables precise dissection of the PGM1-dependent metabolic network.

In the HEK293T background, PGM1 knockout recapitulates key metabolic defects observed in PGM1 deficiency, a congenital disorder of glycosylation (CDG1T) characterized by hypoglycemia, myopathy, cardiomyopathy, and abnormal glycogen accumulation. This cell-based model allows controlled investigation of how loss of phosphoglucomutase activity disrupts glucose-1-phosphate/glucose-6-phosphate equilibrium and its downstream consequences without confounding organismal variables. It is particularly valuable for studying galactose sensitivity, as PGM1 deficiency impairs galactose metabolism, and for screening enzyme replacement or small-molecule therapies aimed at restoring metabolic flux. The interaction between PGM1 and glycogen phosphorylase can be explored in a simplified system, offering insights into glycogen storage disorders and related myopathies.

Research applications of the PGM1 Knockout HEK293T Cell Line encompass a broad spectrum of metabolic and disease-focused studies. The knockout line can be used for quantitative analysis of PGM1 enzymatic activity loss via spectrophotometric assays, metabolic labeling with [13C]glucose to trace carbon flux, and glycogen content measurement. Functional assays include cell viability under glucose deprivation, galactose toxicity challenges, and glucose-6-phosphate quantification. Companion tools such as western blotting and RT-qPCR confirm PGM1 disruption at the protein and transcript levels. This model further supports investigations into insulin and glucagon signaling pathways that regulate PGM1 expression and activity. For detailed product inquiries or technical support, please contact Ascent Research.