The ALDH3A1 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of the near-haploid human HAP1 cell line carrying targeted disruptions in the ALDH3A1 gene. This product provides a mixed pool of knockout cells, enabling loss-of-function studies of aldehyde dehydrogenase 3A1 without clonal selection. The polyclonal format reflects the heterogeneous nature of CRISPR-mediated gene editing, offering a robust model for functional assays in a genetically simplified background.
HAP1 is an adherent, fibroblast-like near-haploid cell line originally derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype facilitates efficient CRISPR/Cas9-mediated gene disruption and high-throughput genetic screening, eliminating interference from wild-type allele compensation. This makes HAP1 an established system for knockout studies, phenotype-driven analyses, and unbiased investigation of gene function in human cellular contexts.
ALDH3A1 encodes a cytoplasmic NAD(P)+-dependent aldehyde dehydrogenase that catalyzes the oxidation of a broad range of cytotoxic aldehydes??including 4-hydroxynonenal, malondialdehyde, and acrolein??to their corresponding carboxylic acids, thereby protecting cellular macromolecules from oxidative and electrophilic damage. Its expression is transcriptionally activated by NRF2 (NFE2L2) binding to antioxidant response elements (AREs) upon KEAP1 inactivation by oxidative stress, with additional regulatory inputs from AHR and PPAR??. The enzyme interacts with NAD(P)+ as a cofactor and, in corneal tissue, associates with ??-Crystallin to maintain transparency and refractive properties. Downstream, ALDH3A1 activity lowers intracellular reactive aldehyde levels, attenuating lipid peroxidation cascades and reducing DNA damage, apoptosis, and lipid peroxidation, and it has been implicated in chemoresistance across multiple cancer types.
In the haploid HAP1 background, disruption of ALDH3A1 provides a clean loss-of-function model for dissecting aldehyde detoxification and stress-response pathways without confounding wild-type allele effects. This system is particularly relevant for investigating ALDH3A1’s roles in corneal biology, where it functions as a crystallin, and in cancer, where its upregulation can contribute to drug resistance. The cells also retain leukemic characteristics, enabling exploration of ALDH3A1’s impact on hematopoietic malignancies and oxidative stress management in a near-haploid leukemic context.
Key research applications include Aldefluor enzymatic activity assays, cell viability and cytotoxicity studies under oxidative challenge with agents such as H2O2 or 4-hydroxynonenal, NRF2/ARE luciferase reporter assays, colony formation assays, Western blotting, RT-qPCR, and immunofluorescence to validate knockout efficiency and downstream signaling alterations. This polyclonal knockout model is also suited for genetic toxicity and chemical screening, chemoresistance profiling, corneal dystrophy and cataract modeling, and engineering cell lines for bioproduction. For further information, please contact Ascent Research.
