The DUSP12 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HEK293T human embryonic kidney cell line. This polyclonal pool provides a loss-of-function model for investigating dual-specificity phosphatase 12 (DUSP12) without single-cell cloning, preserving genetic diversity while ensuring robust DUSP12 ablation suitable for high-throughput screening applications.
HEK293T cells originate from female human embryonic kidney, grow adherently, and stably express the SV40 large T antigen along with integrated adenovirus 5 DNA. These features confer exceptional transfection efficiency and support episomal replication of plasmids containing the SV40 origin of replication, making the line broadly employed for recombinant protein expression, viral production, and transient overexpression studies. Their genetic tractability provides an optimal chassis for CRISPR-mediated gene disruption.
DUSP12 is a dual-specificity phosphatase that dephosphorylates glucokinase (GCK) at key regulatory residues, modulating GCK activity and nuclear-cytoplasmic shuttling in response to metabolic cues including glucose, insulin, and glucagon. It functions within a complex containing GCK and the glucokinase regulatory protein (GKRP) and is regulated by AMPK and endoplasmic reticulum stress. By controlling the conversion of glucose to glucose-6-phosphate, DUSP12 governs hepatic glycolytic flux and integrates signals from the insulin and glucagon receptors.
Although HEK293T cells lack endogenous hepatic factors, their high transfectability permits reconstitution of the DUSP12?CGCK axis through exogenous expression of GCK and other pathway components. The knockout background eliminates endogenous phosphatase activity, enabling precise investigation of GCK phosphorylation, protein?Cprotein interactions, and downstream metabolic effects. The polyclonal format also supports pooled screening strategies to identify novel DUSP12 regulators.
Applications include Western blotting and phospho-specific antibody detection of GCK phosphorylation, co-immunoprecipitation of DUSP12?CGCK complexes, glucose uptake assays, and Seahorse-based glycolysis stress tests. The cells are suitable for RT-qPCR profiling of metabolic genes and immunofluorescence visualization of GCK shuttling. These assays facilitate research into hepatic glucose metabolism, type 2 diabetes, non-alcoholic fatty liver disease, and hepatocellular carcinoma, as well as pharmacological screening of DUSP12 modulators. For further information, contact Ascent Research.
