The HDDC3 Knockout HeLa Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical adenocarcinoma cell line, engineered to disrupt the endogenous HDDC3 gene. This validated polyclonal knockout model enables loss-of-function studies of HDDC3, a cytosolic NADPH phosphatase that regulates cellular redox homeostasis. The polyclonal format provides a genetically heterogeneous population, reflecting the varied editing outcomes typical of CRISPR/Cas9-mediated gene disruption, and is suitable for pooled functional screening or bulk biochemical assays. As a ready-to-use product, these cells facilitate robust investigation of HDDC3-dependent pathways without requiring researchers to perform genome editing from scratch.
HeLa cells are an immortalized epithelial cell line originally isolated from a cervical adenocarcinoma and are among the most widely used human cell lines in biomedical research. Their robust growth characteristics, genetic tractability, and extensive characterization make them a favored model for studying oncogenic signaling, drug response, and fundamental cell biology. The HeLa background provides a well-defined cancer cell context in which to interrogate the function of HDDC3, an enzyme linked to ferroptosis regulation and NADPH metabolism. These cells retain key features of their cancerous origin, including dysregulated proliferation and altered redox balance, which make them particularly suitable for dissecting pathways that intersect oxidative stress and tumor cell survival.
HDDC3 encodes a cytosolic NADPH phosphatase that dephosphorylates NADPH to NADH, thereby reducing cytosolic NADPH levels. This impairs glutathione-dependent antioxidant defense, promoting lipid peroxidation and sensitizing cells to ferroptosis, an iron-dependent cell death pathway. Expression of HDDC3 is activated by ATF4 in response to oxidative or ER stress, positioning it as a stress-responsive regulator of NADPH metabolism. Downstream, decreased NADPH availability compromises GPX4-mediated lipid peroxide reduction, while glutathione metabolism and SLC7A11 are coordinately affected. HDDC3 directly interacts with NADPH as a substrate and may functionally interface with NAD kinases to influence the overall NADP(H) pool.
In the HeLa cervical adenocarcinoma model, HDDC3 knockout provides a powerful tool for investigating the enzyme’s role in ferroptosis regulation. By disrupting the NADPH phosphatase activity, these cells reveal how NADPH dynamics influence lipid peroxidation and ferroptotic cell death, thereby shedding light on redox vulnerabilities in cervical cancer and beyond. This model is particularly suited for pharmacological studies targeting NADPH metabolism or ferroptosis induction.
Typical applications include NADPH/NADP+ quantification, glutathione assays, and lipid peroxidation measurement using C11-BODIPY. Ferroptosis induction with erastin or RSL3 coupled to viability assays evaluates HDDC3-dependent sensitivity. Molecular analysis via western blotting for GPX4 and SLC7A11, and RT-qPCR for HDDC3, confirms knockout and pathway effects. For further information, contact Ascent Research.
