The DNPH1 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the DNPH1 gene in Huh-7 cells. This product consists of a heterogenous pool of edited cells, each carrying distinct loss-of-function alleles, providing a robust loss-of-function model without monoclonal selection. The polyclonal format maintains genetic diversity while abolishing DNPH1 function, suitable for experiments requiring population-level knockout effects.
The Huh-7 cell line, derived from a liver tumor in a 57-year-old Japanese male, is a well-established hepatocellular carcinoma model. It retains key hepatocyte features, including active hepatic metabolism and susceptibility to viral infection, making it widely used in liver cancer research, drug metabolism studies, and host-pathogen interactions. Its ease of culture and genetic manipulability further support its application in CRISPR-based functional genomics.
DNPH1 encodes 6-pyruvoyltetrahydropterin synthase (PTPS), catalyzing the second step of tetrahydrobiopterin (BH4) synthesis. BH4 is a critical cofactor for aromatic amino acid hydroxylases (phenylalanine hydroxylase, tyrosine hydroxylase, tryptophan hydroxylase) and nitric oxide synthases (NOS1, NOS2, NOS3), thereby regulating neurotransmitter production and nitric oxide signaling. PTPS functions downstream of GTP cyclohydrolase I (GCH1) and interacts with sepiapterin reductase (SPR). Its expression is induced by TNF-??, LPS, and interferon-gamma, linking BH4 biosynthesis to inflammation.
In Huh-7 cells, DNPH1 knockout enables investigation of BH4-dependent pathways in hepatocellular carcinoma. Liver is a hub for pterin metabolism; loss of PTPS impairs de novo BH4 production, impacting nitric oxide availability and aromatic amino acid homeostasis. This model can reveal how cancer cells adapt BH4 salvage or alter folate cycle intermediates, affecting proliferation, redox balance, and metabolic reprogramming. It also permits study of tumor-associated nitric oxide effects on angiogenesis and immune modulation.
Typical applications include BH4 quantification by HPLC, PTPS western blotting, RT-qPCR for DNPH1, Griess assay for nitric oxide, neurotransmitter profiling, co-immunoprecipitation with GCH1/SPR, and cell proliferation assays. This knockout cell population is valuable for drug target validation, metabolism studies, and disease modeling of BH4-deficient hyperphenylalaninemia and neurotransmitter disorders. For further information, contact Ascent Research.
