DUSP12 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of HAP1 cells, in which the DUSP12 gene has been disrupted by CRISPR/Cas9-mediated gene editing. This product provides a loss-of-function model for studying DUSP12 in a near-haploid human cell background. The polyclonal population consists of a heterogeneous mix of DUSP12-disrupted cells, enabling the assessment of gene function without clonal selection biases.
The HAP1 cell line is a near-haploid, adherent human cell line derived from the male chronic myeloid leukemia line KBM-7. Due to its haploid karyotype, HAP1 is widely used in haploid genetic screens, drug target discovery, and functional genomics. Its stable, adherent growth and robust proliferation facilitate a broad range of downstream assays, including metabolic flux analysis and western blotting.
DUSP12 encodes an atypical dual-specificity phosphatase that specifically dephosphorylates and activates glucokinase (GCK), a key enzyme that phosphorylates glucose to glucose-6-phosphate in the first step of glycolysis. This activation promotes glucose metabolism and energy production. DUSP12 is regulated by oxidative stress and cellular energy status, and it interacts with 14-3-3 proteins. Downstream of GCK activation, glucose flux influences the insulin receptor and GLUT2 transporter systems. Loss of DUSP12 function may impair GCK activity, leading to reduced glucose phosphorylation, altered cellular energy homeostasis, and increased susceptibility to apoptosis.
In the HAP1 background, DUSP12 knockout provides a powerful system to dissect GCK-dependent and -independent functions of this phosphatase. The near-haploid nature of HAP1 simplifies the interpretation of gene-disruption phenotypes, as a single copy of the gene is targeted. Moreover, the chronic myeloid leukemia origin of HAP1 links this model to cancer metabolism research, where altered glucose metabolism is a hallmark. The polyclonal knockout population allows investigators to study the collective impact of DUSP12 loss while mitigating clonal artifacts, making it suitable for drug target validation in metabolic syndrome, type 2 diabetes, and cancer.
Typical applications include investigation of glucose metabolism regulation via glucose uptake assays and GCK activity measurements, study of post-translational control of GCK by DUSP12 using western blotting and RT-qPCR, metabolic disease modeling, and apoptosis assays with caspase-3/7 detection. Additionally, this knockout can be employed in functional genomics screens to identify synthetic lethal interactions or modulators of cellular stress responses, with readouts such as cell viability and metabolic flux analysis. For more information on product specifications and availability, please contact Ascent Research.
