The DNPH1 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma line, designed for loss-of-function analysis of the DNPH1 gene. This heterogeneous knockout pool arises from targeted gene disruption, enabling robust functional studies without clonal selection artifacts.
The parental HT29 cell line was isolated from a colorectal adenocarcinoma of a 44-year-old female and serves as an established epithelial model for colonic cancer biology and intestinal barrier function. These adherent cells maintain key hallmarks of colorectal tumorigenesis, including oncogenic signaling and metabolic rewiring, offering a relevant context for examining nucleotide metabolism and mitochondrial dynamics.
DNPH1 encodes a nucleoside monophosphate hydrolase that catalyzes the hydrolysis of deoxynucleoside monophosphates into deoxynucleosides, a central step in nucleotide salvage and the regulation of mitochondrial deoxynucleotide precursor pools. Transcription of DNPH1 is driven by the c-Myc oncoprotein, placing it downstream of major proliferative signals. The hydrolytic activity of DNPH1 ultimately modulates deoxynucleoside availability and mitochondrial DNA synthesis, with substrate flux feeding nucleotide kinases and mitochondrial DNA polymerase gamma.
In the HT29 colorectal cancer background, knockout of DNPH1 disrupts nucleotide homeostasis, potentially compromising mitochondrial DNA replication and cell proliferation. This polyclonal knockout population therefore provides a physiologically relevant system to investigate how altered nucleotide metabolism influences genomic instability, metabolic reprogramming, and tumor cell fitness. It also offers a platform for studying mitochondrial DNA depletion mechanisms in an epithelial cancer setting.
Typical research applications include quantitative nucleotide profiling by LC-MS, mitochondrial DNA copy number assessment by qPCR, cell proliferation and ATP measurement, and chemosensitivity screening with nucleoside analogs or nucleotide?targeting compounds. Validation of DNPH1 disruption can be performed via western blotting or RT-qPCR. For additional information or technical support, please contact Ascent Research.
