HPRT1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human lung adenocarcinoma NCI-H1975 cell line, featuring targeted disruption of the HPRT1 gene. This polyclonal format offers a heterogeneous collection of edited alleles, providing a robust loss-of-function model that captures phenotypic variability and avoids the biases of clonal isolation. It is well-suited for investigating purine salvage pathway depletion in a cancer-relevant epithelial background.
The parental NCI-H1975 cell line originates from a female patient with non-small cell lung cancer (NSCLC) and is widely utilized for studying oncogenic signaling, drug sensitivity, and metabolic reprogramming. These adherent cells maintain an epithelial morphology and key genetic alterations, making them a representative platform for lung adenocarcinoma research. Introduction of HPRT1 knockout into this context enables the examination of nucleotide metabolism in NSCLC.
HPRT1 encodes hypoxanthine-guanine phosphoribosyltransferase, an enzyme that catalyzes the salvage synthesis of inosine monophosphate (IMP) and guanosine monophosphate (GMP) from hypoxanthine and guanine, respectively, using phosphoribosyl pyrophosphate (PRPP) as a co-substrate. Transcription of HPRT1 is regulated by Sp1 and Myc, and its activity depends on intracellular PRPP levels and purine availability. As a central node in the purine salvage pathway, HPRT1 acts upstream of nucleotide interconversion, with IMP and GMP feeding into pools essential for DNA and RNA synthesis. Core pathway components include PRPS1, APRT, ADA, PNP, IMPDH, GMPS, and XDH. CRISPR/Cas9-mediated knockout abrogates salvage activity, leading to accumulation of hypoxanthine and guanine, depletion of IMP and GMP, and induction of de novo purine biosynthesis. This metabolic shift confers resistance to purine analog prodrugs such as 6-thioguanine and 6-mercaptopurine, which require HPRT1 for bioactivation.
In the context of NCI-H1975 lung adenocarcinoma, HPRT1 knockout forces reliance on de novo purine synthesis, exposing potential metabolic vulnerabilities. The model is therefore valuable for studying drug resistance mechanisms, particularly to purine antimetabolites, and for screening synthetic lethal partners with inhibitors targeting de novo pathway enzymes. Moreover, the accumulation of purine metabolites offers a system for investigating hyperuricemia-related pathologies and gout.
This polyclonal knockout product is suitable for a range of research applications, including purine metabolism studies, Lesch-Nyhan disease modeling, gene targeting selection strategies, and drug resistance screening in NSCLC. Standard analytical techniques such as Western blotting, RT-qPCR, and Sanger sequencing can be used to confirm knockout at the protein, mRNA, and genomic levels. Functional assays like HPRT enzymatic activity measurement and 6-thioguanine resistance testing directly evaluate loss of function, while metabolomic analyses via LC-MS or HPLC enable detailed profiling of nucleotide disruptions. For further inquiries, please contact Ascent Research.
