ADH5 Knockout A-549 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma epithelial cell line, with targeted disruption of the ADH5 gene. This loss-of-function polyclonal model enables functional studies of alcohol dehydrogenase 5 (formaldehyde dehydrogenase) in a physiologically relevant pulmonary epithelial context. The product is provided as a viable polyclonal stock suitable for expansion and applications requiring stable gene ablation without clonal selection.
The A-549 host cell line, originally isolated from lung carcinoma tissue of a 58-year-old Caucasian male, is widely used to model alveolar epithelial cell biology, surfactant production, and xenobiotic metabolism. Exhibiting type II pneumocyte features, these adherent cells are employed in drug transport, metabolic enzyme expression, and respiratory toxicology studies. Their robust growth and well-characterized transcriptome offer an ideal platform for ADH5 knockout derivatives to investigate formaldehyde-induced stress responses.
ADH5 encodes a glutathione-dependent formaldehyde dehydrogenase that catalyzes oxidation of S-hydroxymethylglutathione to S-formylglutathione, a critical step in formaldehyde detoxification and glutathione cycling. The enzyme is transcriptionally regulated by NRF2 (NFE2L2) via oxidative stress response elements, linking ADH5 expression to cellular redox homeostasis. ADH5 interacts with glutathione, formaldehyde, and zinc ion as cofactor. In the knockout model, formaldehyde accumulation and S-hydroxymethylglutathione build-up lead to increased DNA adducts and genotoxic stress. Downstream, formylglutathione hydrolase converts S-formylglutathione to glutathione and formate; impaired clearance enhances ??H2AX phosphorylation and activates apoptotic cascades.
Abolishing ADH5 in A-549 cells recapitulates key features of formaldehyde toxicity syndromes and cancer predisposition observed in bone marrow failure and leukemia. ADH5 deficiency disrupts neutralization of endogenous and exogenous formaldehyde, causing DNA interstrand crosslinks, replication stress, and impaired proliferation. This model enables dissection of genotoxic stress responses specifically within a lung adenocarcinoma background, where environmental formaldehyde exposure is a recognized risk factor. The polyclonal population facilitates study of how detoxification loss influences epithelial-to-mesenchymal transition, apoptosis, and synthetic lethal interactions in the tumor microenvironment.
Researchers can employ this knockout population for formaldehyde toxicity assays using cell viability or apoptosis readouts, DNA damage and repair studies via comet assay or ??H2AX immunofluorescence, and enzymatic activity measurements to confirm ADH5 ablation. Transcriptomic profiling (RNA-seq) can map global stress responses, while the model also supports cancer metabolism research, drug toxicity screening, and tumor microenvironment investigations where formaldehyde detoxification intersects with metabolic reprogramming. For ordering or technical inquiries, please contact Ascent Research.
