The ACADM Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma line, featuring disrupted ACADM expression to abolish medium-chain acyl-CoA dehydrogenase activity. This heterogeneous pool serves as a loss-of-function model for studying medium-chain fatty acid oxidation defects in a well-characterized epithelial background, avoiding clonal artifacts.
The A-549 cell line, established from a lung carcinoma patient, is a widely used model of human alveolar type II epithelium with adherent morphology. It is extensively applied in cancer research, drug metabolism, and toxicology studies due to its robust growth characteristics and relevance to pulmonary biology, making it an ideal host for metabolic pathway interrogation.
ACADM encodes a mitochondrial matrix enzyme that catalyzes the initial step of ??-oxidation for medium-chain (C4?CC12) fatty acids, converting acyl-CoAs to trans-2-enoyl-CoA and delivering electrons to the electron transfer flavoprotein (ETF) complex. ACADM interacts directly with ETFA and ETFB, and its activity is functionally coupled to ETFDH. Transcriptional control is exerted by PPARA, NR1H4, HNF4A, and SIRT1, while insulin/glucagon signaling further modulates expression. Downstream, ACADM activity generates acetyl-CoA, NADH, and FADH2, positioning it at a critical node linking lipid catabolism to energy homeostasis and signaling. This pathway is integrated with the mitochondrial trifunctional protein (HADHA/HADHB) and other acyl-CoA dehydrogenases (ACADVL, ACADS).
In A-549 cells, ACADM knockout impairs medium-chain fatty acid utilization, potentially redirecting metabolism toward glycolysis and altering nutrient dependencies. This provides a controllable system to model medium-chain acyl-CoA dehydrogenase deficiency (MCADD) and to study the impact of fatty acid oxidation defects on cancer cell bioenergetics and survival. Moreover, the knockout enables dissection of the PPARA?CSIRT1 regulatory axis in a tumorigenic environment.
Applications include Seahorse metabolic flux analysis, 14C-palmitate oxidation assays, targeted acyl-carnitine profiling, and glucose-free survival experiments. The cells are also suited for cell proliferation assays, immunofluorescence-based mitochondrial morphology studies, and drug metabolism screening; they are particularly valuable for investigating metabolic reprogramming in cancer and for screening modulators of fatty acid oxidation. Knockout can be verified by Western blotting and RT-qPCR. For further inquiries, please contact Ascent Research.
