The ALDH2 Knockout HAP1 Polyclonal Cells comprise a population of CRISPR/Cas9-edited polyclonal knockout cells generated from the HAP1 cell line, with targeted disruption of the human ALDH2 gene. This product provides a genetically mixed pool of cells harboring CRISPR/Cas9-mediated gene disruption, enabling loss-of-function studies without clonal selection. It serves as a versatile tool for researchers investigating aldehyde metabolism and related pathologies.
HAP1 is a human near-haploid fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia line. It exhibits both adherent and suspension growth characteristics and retains a stable male karyotype with haploid conversion. Its near-haploid nature reduces genetic redundancy, making it a preferred model for knockout studies by facilitating unambiguous genotype-phenotype correlations. The HAP1 background supports robust endogenous expression of metabolic enzymes, providing a relevant context for examining mitochondrial and cytosolic pathways.
ALDH2 encodes mitochondrial aldehyde dehydrogenase 2, which oxidizes acetaldehyde to acetate and participates in retinoic acid biosynthesis from retinaldehyde. Its activity is regulated by upstream factors including ethanol exposure, oxidative stress, and the transcription factors NRF2 and PPARalpha, while retinoic acid itself modulates ALDH2 expression. Downstream, ALDH2-mediated catalysis produces NADH and acetate, contributing to acetaldehyde clearance and retinoic acid signaling. The enzyme functions in concert with interacting partners such as ALDH1B1, the cofactor NAD+, ethanol-metabolizing CYP2E1, and the mitochondrial import receptor TOMM20, within pathways that also include alcohol dehydrogenase 1B (ADH1B) and catalase.
Disruption of ALDH2 in HAP1 cells eliminates mitochondrial aldehyde dehydrogenase activity, impairing acetaldehyde detoxification and causing intracellular accumulation of toxic aldehydes. This leads to elevated oxidative stress and abrogated retinoic acid signaling, mimicking conditions relevant to alcohol intolerance, esophageal cancer, hypertension, and neurodegenerative disorders such as Alzheimer??s and Parkinson’s diseases. The near-haploid background accentuates the functional consequences of ALDH2 loss, facilitating dissection of its role in aldehyde-induced cytotoxicity and mitochondrial dysfunction without interference from a second functional allele.
These polyclonal knockout cells are suited for a broad spectrum of experimental applications, including studies of ethanol metabolism, aldehyde toxicity, oxidative stress responses, and alcohol-related cancer biology. Researchers can employ western blotting to confirm ALDH2 protein loss, enzymatic activity assays for functional validation, and cellular aldehyde quantification to measure substrate accumulation. Additional characterization may involve MTT assays for viability, ROS detection for oxidative stress, RT-qPCR for transcript-level assessment, RNA-seq for global transcriptomic profiling, and comet assays for DNA damage. The polyclonal format offers a heterogeneous population that captures a range of editing outcomes, enabling robust population-level analyses. For further information, please contact Ascent Research.
