The HK1 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population generated by targeted disruption of the human HK1 gene within the HAP1 cell line. This polyclonal pool enables robust analysis of hexokinase 1 deficiency without clonal selection, allowing direct investigation of the metabolic consequences of abrogating the first committed step of glycolysis.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia background, displaying adherent fibroblast-like morphology. Its haploid genome facilitates efficient gene targeting and unambiguous genotype?Cphenotype correlations, making it a premier tool for genetic screening and knockout cell line generation. The leukemic origin also provides a relevant context for studying metabolic reprogramming in haematological malignancies.
HK1 encodes hexokinase 1, which catalyzes the ATP-dependent phosphorylation of glucose to glucose-6-phosphate, the rate-limiting step of glycolysis. The enzyme additionally binds the mitochondrial voltage-dependent anion channel (VDAC) and the adenine nucleotide translocator, coupling glycolytic flux to oxidative phosphorylation and regulating apoptosis through direct interactions with Bax and Bak. HK1 is transcriptionally activated by HIF-1?? and c-Myc and is modulated downstream of the insulin/PI3K/AKT pathway. Its product, glucose-6-phosphate, feeds into glycolysis, the pentose phosphate pathway, and ATP synthesis, where it is further metabolized by glucose-6-phosphate isomerase and phosphofructokinase, thereby integrating multiple metabolic and signaling networks.
Disruption of HK1 in HAP1 creates a metabolic bottleneck that uncouples glycolysis from mitochondrial respiration and may sensitize cells to apoptotic triggers. The near-haploid state ensures that phenotypic outcomes are directly attributable to HK1 loss, providing a clean system for dissecting metabolic control points relevant to cancer proliferation, insulin action, and disorders such as hereditary hexokinase deficiency and hemolytic anemia.
Applications span cancer metabolism screens, glycolysis inhibitor testing, diabetes research on insulin-stimulated glucose utilization, and apoptosis regulation studies. Downstream assays include western blotting for HK1 validation, hexokinase activity and glucose uptake measurements, lactate production and mitochondrial membrane potential assessments, Annexin V apoptosis detection, and co-immunoprecipitation with VDAC. For technical inquiries, please contact Ascent Research.
