The HDHD2 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population targeting the HDHD2 gene in the HAP1 cell line. This heterogeneous pool of cells harboring targeted gene disruptions serves as a robust loss-of-function model, avoiding clonal variation while enabling reliable functional analyses of HDHD2-dependent processes.
HAP1 is a near-haploid human cell line derived from the chronic myeloid leukemia KBM-7 line, originally from a male patient. Adapted for suspension culture, HAP1 maintains near-haploidy except for chromosome 8 disomy, reducing genetic redundancy and simplifying CRISPR-based genetic screens. Its CML origin also provides a relevant background for cancer metabolism studies.
HDHD2 belongs to the HAD superfamily of hydrolases and functions as a putative pseudouridine-5??-phosphatase. According to current mechanistic understanding, HDHD2 dephosphorylates pseudouridine-5??-phosphate to pseudouridine, a key step in pseudouridine degradation and nucleotide salvage. Upstream, pseudouridine kinase phosphorylates pseudouridine to generate the substrate, while HDHD2 acts downstream to recycle the nucleoside. Representative pathway components include pseudouridine, pseudouridine-5??-phosphate, pseudouridine kinase, and HDHD2, placing this enzyme at the intersection of pyrimidine metabolism and salvage pathways.
The combination of HDHD2 knockout with HAP1??s haploid genome creates an efficient platform for dissecting nucleotide metabolism. CRISPR/Cas9 disruption in a near-haploid background often leads to complete loss-of-function, enabling clear assessment of pseudouridine catabolism impacts on pyrimidine pools and salvage efficiency. This polyclonal population is particularly suited for high-throughput screens and metabolic flux analyses requiring uniform genetic backgrounds.
Typical applications include functional genomics, nucleotide metabolism studies, and exploration of metabolic disease mechanisms. Researchers can confirm HDHD2 disruption via RT-qPCR and western blot, measure phosphatase activity biochemically, and perform LC-MS-based nucleotide profiling. Cell viability assays can also assess metabolic vulnerabilities. This product serves as a versatile tool for both targeted pathway analysis and broader phenotypic screens. For further details or technical support, please contact Ascent Research.
