The ADCK2 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HAP1 human near-haploid cells with disrupted ADCK2, an atypical mitochondrial protein kinase critical for coenzyme Q (CoQ) biosynthesis. This loss-of-function model is designed to impair ADCK2 function, enabling researchers to study mitochondrial energy metabolism and CoQ-deficiency mechanisms without clonal selection bias.
HAP1 is a male chronic myeloid leukemia-derived adherent cell line with a near-haploid karyotype and disomy of chromosome 8, prized for its efficiency in haploid genetic screens and gene-editing applications. The reduced genomic redundancy simplifies phenotypic interpretation and enhances the ability to detect subtle metabolic defects, making it an ideal host for interrogating CoQ pathway function.
ADCK2 is a mitochondrial kinase that phosphorylates COQ3, COQ5, COQ7, and PDSS1 to promote CoQ biosynthesis, which is essential for electron transfer between respiratory complexes I?CIII. Its expression is induced by PPARGC1A and modulated by AMPK and mTOR, integrating metabolic signals. ADCK2 interacts with COQ3, COQ5, COQ7, ADCk3, PDSS1, and PDSS2, channeling substrates into the electron transport chain (Complexes I?CIV and ATP synthase). Disruption of ADCK2 lowers CoQ levels, impairing oxidative phosphorylation and ATP synthesis, and recapitulates molecular defects observed in primary CoQ10 deficiency and mitochondrial encephalomyopathies.
In the HAP1 background, ADCK2 knockout provides an unambiguous human cellular model for dissecting CoQ deficiency phenotypes, including those associated with nephrotic syndrome and mitochondrial disorders. The haploid system allows direct assessment of mitochondrial respiration by Seahorse respirometry, CoQ10 quantification via HPLC, and immunofluorescence staining of mitochondrial morphology without interference from wild-type alleles. Paired with parental HAP1 cells, this polyclonal model enables robust comparative analyses of pathway compensation and metabolic adaptation.
Applications include ATP bioluminescence assays for bioenergetics profiling, RT-qPCR screening of CoQ pathway genes, and CRISPR-based genetic screens to identify modifiers of ADCK2 loss. Drug discovery can deploy these cells in high-throughput screens to rescue CoQ levels or restore oxidative phosphorylation, with readouts from HPLC and respirometry. Western blotting and co-immunoprecipitation validate ADCK2-dependent phosphorylation of COQ3/COQ5/COQ7, while metabolic flux assays explore shifts in glycolysis and fatty acid oxidation. For further information or to place an order, please contact Ascent Research.
