The AANAT Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from human A-549 alveolar basal epithelial cells, engineered for loss-of-function analyses of the arylalkylamine N-acetyltransferase (AANAT) gene. The polyclonal composition ensures allelic diversity, avoiding the confounding effects of clonal selection and providing a robust platform for functional genomic studies without introducing single-cell bottleneck artefacts.
A-549 cells, originally isolated from a lung adenocarcinoma resected from a 58-year-old male, represent a well-characterized model of type II pulmonary epithelial cells. Extensively employed in respiratory oncology, epithelial barrier biology, and viral infection research, these cells offer a genetically tractable background for dissecting oncogenic pathways and cancer cell-autonomous metabolic processes.
AANAT catalyzes the rate-limiting acetylation of serotonin to N-acetylserotonin in melatonin biosynthesis. Its expression is circadian-regulated by noradrenaline activating ??-adrenergic receptors (e.g., ADRB1) and downstream cAMP?CPKA?CCREB signaling, with transcriptional input from the BMAL1/CLOCK complex. Stability is enhanced by 14-3-3 protein binding. Downstream, N-acetylserotonin is methylated by ASMT to melatonin, which engages MTNR1A/B receptors and upregulates antioxidant enzymes including GPx and SOD.
In A-549 cells, AANAT abrogation disrupts autocrine melatonin production, permitting distinction of intracellular from paracrine melatonin effects in lung cancer. This model addresses how loss of circadian melatonin synthesis influences proliferation, redox homeostasis, and metastatic potential, relevant to the emerging role of melatonin in cancer suppression and circadian disruption.
Applications include RT-qPCR and western blotting for knockout confirmation, melatonin quantification by ELISA or LC-MS/MS, and functional assays such as MTT viability, ROS detection, and cell migration studies. These cells further support circadian bioluminescence reporter assays and high-throughput drug screening for melatonin pathway modulators. For additional technical specifications or ordering, please contact Ascent Research.
