The DNPH1 Knockout UM-UC-3 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population derived from the UM-UC-3 bladder cancer line, carrying a disrupted DNPH1 gene. This polyclonal pool avoids clonal bias and preserves heterogeneous mutations, making it ideal for studying DNPH1 loss-of-function effects. The cells are generated by CRISPR/Cas9-mediated gene disruption and are suited for applications in nucleotide metabolism, DNA damage, and cancer therapeutic research.
UM-UC-3 is a human male urinary bladder transitional cell carcinoma line with mutant TP53 and null PTEN. These defects impair tumor suppression, leading to inherent genomic instability and altered DNA damage responses. Thus, UM-UC-3 provides a sensitized background for investigating pathways maintaining genome integrity, such as those involving DNPH1.
DNPH1 functions as a dNTP pyrophosphohydrolase that sanitizes nucleotide pools by hydrolyzing noncanonical dNTPs, notably dUTP, preventing their incorporation into DNA. Loss of DNPH1 causes accumulation of aberrant dNTPs, an elevated dUTP/dTTP ratio, and uracil misincorporation. This triggers futile repair cycles by uracil-DNA glycosylase (UNG) and AP endonuclease, resulting in DNA breaks and genomic instability. DNPH1 is transcriptionally controlled by E2F factors and DNA damage signaling, connecting nucleotide pool quality to replication fork integrity.
In UM-UC-3 cells lacking TP53 and PTEN, DNPH1 knockout exacerbates replication stress and DNA damage, potentially creating synthetic vulnerabilities. This may alter reliance on repair pathways or enhance sensitivity to DNA-damaging agents, offering a platform to explore interactions between nucleotide pool dysregulation and oncogenic signaling in bladder cancer.
These cells support diverse investigations, including cancer biology, DNA damage response, and nucleotide metabolism. Key assays include Western blotting for DNPH1 and ??H2AX, dNTP pool quantification, comet assay, clonogenic survival, and drug sensitivity profiling. For further details, contact Ascent Research.
