This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, featuring targeted disruption of the HDDC2 gene. Using CRISPR/Cas9-mediated gene disruption, this knockout model provides a loss-of-function tool to investigate HDDC2’s role in nucleotide metabolism and antiviral defense. The polyclonal format maintains population heterogeneity while effectively abrogating HDDC2 expression, suitable for pooled functional studies.
HeLa cells are an immortalized human cervical adenocarcinoma epithelial cell line, originally isolated from a patient with cervical cancer. They harbor human papillomavirus type 18 (HPV-18) sequences, resulting in inactivation of the p53 and retinoblastoma (Rb) tumor suppressor pathways. This genetic background drives rapid proliferation and has established HeLa as a widely used model in cancer biology, virology, and cell signaling research.
HDDC2 encodes a putative deoxynucleoside triphosphate (dNTP) triphosphohydrolase that hydrolyzes dNTPs, thereby lowering intracellular dNTP pools and restricting DNA synthesis and viral replication. Its expression is transcriptionally regulated by interferon alpha/beta signaling through STAT1 and IRF1, linking it to innate immunity. Key downstream effects include modulation of dNTP concentrations, DNA polymerase activity, and viral replication efficiency. Representative pathway components include ribonucleotide reductase subunits RRM1 and RRM2, thymidine kinase TK1, interferon receptor IFNAR1, and STAT1. Although direct interacting partners are uncharacterized, HDDC2 likely associates with nucleotide metabolism enzymes.
In HeLa cells, which exhibit deregulated cell cycle control and high nucleotide demand, HDDC2 depletion may elevate dNTP pools, potentially enhancing DNA replication and proliferation. This model is particularly valuable for dissecting the interplay between nucleotide metabolism and oncogenic transformation, as well as for examining how interferon-induced restriction factors limit viral propagation in HPV-positive cervical cancer cells. Given HDDC2’s homology to SAMHD1, which is implicated in autoimmune disorders, this knockout system may also aid in exploring uncharacterized links to autoimmunity.
The HDDC2 knockout HeLa polyclonal cells are suited for functional characterization of nucleotide metabolism, employing assays such as dNTP pool measurement by HPLC, RT-qPCR for HDDC2 mRNA quantitation, and western blotting for protein analysis. Proliferation and cell cycle effects can be assessed via EdU incorporation and flow cytometry, while antiviral roles are probed using viral infectivity assays. These cells provide a robust platform for innate immunity pathway analysis and cancer cell proliferation studies. For further details, please contact Ascent Research.
