The HDDC3 Knockout HEK293T Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population in which the human HDDC3 gene has been disrupted to generate a loss-of-function model. This polyclonal format provides a heterogeneous pool of edited cells, enabling robust functional studies without clonal selection artifacts. The knockout eliminates HDDC3 expression, abrogating its native enzymatic activity and allowing researchers to dissect its role in cellular redox regulation and ferroptosis sensitivity.
The host cell line, HEK293T, is an extensively characterized human embryonic kidney cell line transformed with SV40 large T antigen. HEK293T cells are renowned for their high transfection efficiency, rapid growth, and exceptional capacity for recombinant protein expression and lentiviral production. Their robust metabolic machinery and well-defined signaling networks make them an ideal chassis for investigating gene function, particularly in pathways governing stress responses and cell death mechanisms.
HDDC3 encodes a NADPH phosphatase that dephosphorylates NADPH to NADH, lowering the NADPH/NADP+ ratio and sensitizing cells to ferroptosis, a lipid peroxidation-driven necrosis. It interacts with ferroptosis regulators and NADPH-consuming enzymes, and its activity is modulated by nutrient stress and redox imbalance. Loss of HDDC3 elevates NADPH, conferring resistance to ferroptosis inducers like Erastin and RSL3, and attenuates engagement of downstream effectors GPX4, SLC7A11, and ACSL4. Additionally, HDDC3 may serve as a ppGpp hydrolase during stringent response.
In HEK293T cells, HDDC3 knockout provides a clean system to dissect NADPH homeostasis independent of other cell-type-specific redox mechanisms. Given HEK293T’s utility in protein expression and viral production, this model enables systematic exploration of how NADPH dynamics affect cellular productivity and stress resilience. It is particularly valuable for studying crosstalk between nucleotide metabolism and ferroptosis in a tractable, high-throughput-compatible background.
Researchers can employ these cells in ferroptosis induction assays (Erastin, RSL3), lipid peroxidation monitoring (C11-BODIPY), NADPH/NADP+ quantification, and ROS flow cytometry. They support western blotting, RT-qPCR, and metabolomic analysis of redox pathways. Applications include ferroptosis studies, cancer research, metabolic disease modeling, and drug sensitivity screening. For further information, contact Ascent Research.
