The ALDH1A3 Knockout A-549 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A-549 cell line, with disruption of the ALDH1A3 gene. ALDH1A3 encodes aldehyde dehydrogenase 1 family member A3, a key enzyme in retinoid metabolism. This polyclonal population features gene disruptions that collectively ablate ALDH1A3 function, providing a robust loss-of-function system for investigating ALDH1A3-dependent processes.
The A-549 host cell line originates from a lung adenocarcinoma of a 58-year-old Caucasian male and exhibits adherent epithelial morphology, serving as a model for type II alveolar epithelial cells and non?small?cell lung cancer (NSCLC). Widely used in cancer biology, A-549 cells express basal ALDH1A3, which contributes to retinoic acid (RA) biosynthesis and is implicated in tumor cell proliferation and stem-like properties. Disrupting ALDH1A3 in this background permits dissection of its oncogenic and differentiation functions.
ALDH1A3 catalyzes the irreversible oxidation of retinaldehyde to all-trans-retinoic acid (ATRA), requiring NAD+ as a cofactor, and is a critical enzyme in RA biosynthesis. ATRA acts as a ligand for nuclear receptors RAR and RXR, which transcriptionally regulate target genes including HOX clusters, CYP26A1, and stemness markers such as NANOG, OCT4, and SOX2. ALDH1A3 expression is controlled by upstream regulators PAX6, SOX2, OTX2, and Wnt signaling. Within the retinaldehyde?CRABP?RA?RAR/RXR axis, ALDH1A3 functions as a rate?limiting step, governing RA availability and downstream transcriptional programs linked to development, differentiation, and cancer.
CRISPR/Cas9-mediated disruption of ALDH1A3 in A-549 cells attenuates retinaldehyde-to-RA conversion, resulting in diminished RA signaling. This loss impairs RA-responsive differentiation programs and alters the proliferation?stemness balance. Consequently, these ALDH1A3 knockout polyclonal cells enable detailed analysis of how RA metabolism shapes lung cancer behavior, including growth kinetics, sphere formation, stem cell marker expression (e.g., CD44, CD133), and responses to differentiation agents.
This knockout model is suited for studying retinoic acid signaling in lung cancer, cancer stem cell biology, differentiation therapy, and drug resistance mechanisms. Compatible assays include RT?qPCR and western blotting for ALDH1A3, HPLC quantification of intracellular RA, flow cytometry for CD44 and CD133, MTS proliferation assays, sphere formation, Boyden chamber migration, and RNA?seq transcriptomics. Comparative analysis with parental A-549 cells allows robust evaluation of ALDH1A3 loss. For further information or technical support, contact Ascent Research.
