The DNPH1 Knockout HEK293T Polyclonal Cells product provides a polyclonal population of HEK293T cells with targeted disruption of the DNPH1 gene, generated using CRISPR/Cas9 technology. This loss-of-function model enables investigation of DNPH1-dependent nucleotide metabolism and c-Myc-mediated pathways without interference from wild-type gene activity, supporting studies in cancer biology, chemoresistance, and nucleotide homeostasis.
HEK293T cells are a widely used human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen, facilitating episomal replication of plasmids bearing the SV40 origin of replication. Derived from the original HEK293 line transformed with sheared adenovirus 5 DNA, these cells are highly transfectable and support robust recombinant protein expression, making them an ideal host for loss-of-function studies interrogating nucleotide metabolism, cell signaling, and drug response mechanisms.
DNPH1 (also known as RCL) is a c-Myc-regulated hydrolase that catalyzes the dephosphorylation of deoxynucleoside 5??-monophosphates, including deoxycytidine monophosphate (dCMP), deoxyuridine monophosphate (dUMP), and deoxythymidine monophosphate (dTMP), to produce deoxycytidine, deoxyuridine, and thymidine, respectively. This enzymatic activity, situated downstream of c-Myc transcriptional regulation, modulates intracellular nucleotide pools and influences DNA synthesis and repair. Elevated DNPH1 expression, driven by c-Myc, is associated with sustained cell proliferation and may reduce sensitivity to nucleoside analog chemotherapeutics.
In the HEK293T background, disruption of DNPH1 eliminates a key node linking c-Myc transcriptional activity to nucleotide homeostasis, providing a defined cellular model to study how altered deoxynucleoside production affects cell cycle progression, DNA damage responses, and chemosensitivity. The polyclonal knockout population preserves cellular heterogeneity, enabling robust and reproducible evaluation of phenotype penetrance across a diverse genetic background without clonal selection artifacts.
Researchers can employ this knockout model for a wide range of applications, including quantitative analysis of nucleotide pools by LC-MS/MS, functional assessment of the c-Myc?CDNPH1 axis using RT-qPCR and western blotting, and systematic drug sensitivity screens with nucleoside analogs such as gemcitabine and cytarabine. The polyclonal knockout cells are also suitable for proliferation and clonogenic assays, flow cytometry-based cell cycle profiling, transcriptomic analyses via RNA-seq, and interaction studies using co-immunoprecipitation. For more information, please contact Ascent Research.
