The HDHD5 Knockout 769-P Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 769-P renal proximal tubule epithelial cell line. This product provides a heterogeneous pool of cells with targeted disruption of the HDHD5 gene, enabling robust loss-of-function studies without clonal selection artifacts. The polyclonal format captures diverse edited alleles, offering a representative model for assessing the biological roles of the encoded haloacid dehalogenase-like phosphatase in a clear cell renal cell carcinoma (ccRCC) background.
The 769-P cell line was established from a primary clear cell renal cell carcinoma of a female patient and is widely used as a model of ccRCC. Originating from the renal proximal tubule epithelium, these cells retain key metabolic and signaling features of the tumor microenvironment, including a strong dependence on nucleotide metabolism for sustained proliferation. This genetic background is particularly relevant for investigating how phosphatase-mediated nucleotide homeostasis intersects with oncogenic processes.
HDHD5 functions as a nucleotide monophosphate phosphatase, catalyzing the dephosphorylation of substrates such as UMP, CMP, and dTMP to regulate intracellular nucleotide pools. Its activity is controlled by substrate availability and transcriptional regulation, and it acts upstream of DNA and RNA synthesis by ensuring a balanced supply of nucleotide precursors. HDHD5 operates within a broader metabolic network that includes CTP synthase, thymidylate synthase, 5′-nucleotidases, and IMP dehydrogenase, collectively coordinating pyrimidine and purine metabolism. Through this network, HDHD5 maintains nucleotide homeostasis critical for genome replication and ribosomal biogenesis in proliferating cells.
In 769-P clear cell renal carcinoma cells, dysregulated nucleotide metabolism fuels unchecked proliferation and tumor progression. Disruption of HDHD5 in this polyclonal knockout population permits systematic interrogation of how loss of this phosphatase perturbs nucleotide pools, nucleic acid synthesis, and cellular energy balance. Given ccRCC??s reliance on de novo nucleotide synthesis, HDHD5 knockout cells provide a relevant model for identifying metabolic vulnerabilities and potential therapeutic targets. Additionally, since HDHD5 has been implicated in craniosynostosis, this model may inform studies of nucleotide imbalance-related developmental disorders when combined with appropriate cellular contexts.
Researchers can employ this polyclonal knockout model to dissect HDHD5 function using phosphatase activity assays and nucleotide quantification by LC-MS, directly measuring changes in monophosphate nucleotide levels. Western blotting and RT-qPCR enable confirmation of gene disruption at the protein and transcript levels, while cell proliferation assays reveal functional consequences on ccRCC growth. These cells are also suitable for craniosynostosis disease modeling by introducing additional genetic or chemical perturbations to recapitulate nucleotide imbalance phenotypes. For further information or technical support, please contact Ascent Research.
