The ACACB Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human A-549 lung adenocarcinoma line. They carry a targeted disruption of the ACACB gene, which encodes acetyl-CoA carboxylase 2 (ACC2), a key enzyme controlling mitochondrial fatty acid oxidation. This polyclonal pool preserves population heterogeneity and eliminates the need for single-cell cloning, providing a convenient loss-of-function model for metabolic research.
The parental A-549 cell line is a hypotriploid, adherent epithelial model of human alveolar Type II pulmonary epithelium, originally isolated from a 58-year-old Caucasian male with lung adenocarcinoma. It is extensively employed in cancer biology, drug metabolism, and virology studies, and its tumorigenic origin makes it particularly relevant for investigating metabolic reprogramming in non-small cell lung cancer.
ACACB encodes ACC2, which converts acetyl-CoA to malonyl-CoA at mitochondria. Malonyl-CoA inhibits CPT1, limiting mitochondrial fatty acid uptake and ??-oxidation. ACC2 is inhibited by AMPK phosphorylation and regulated by PGC-1??, SREBP-1c, and insulin. It interacts with biotin, citrate, palmitoyl-CoA, and MIG12. Knockout eliminates ACC2, lowering malonyl-CoA and disinhibiting CPT1, thus promoting fatty acid oxidation. Key pathway nodes include ACACB, malonyl-CoA, CPT1A, AMPK, and PGC-1??.
In A-549 cells, ACACB disruption provides a model to study how malonyl-CoA signaling controls lipid metabolism in lung adenocarcinoma. Tumor cells often rewire fatty acid oxidation and lipogenesis, and the loss of ACC2 can reveal metabolic vulnerabilities or adaptive responses. By modulating the ACC2?CCPT1A axis, this knockout helps dissect energy homeostasis and its impact on cancer cell proliferation and survival.
Typical applications include metabolic flux analysis with Seahorse, radiolabeled palmitate oxidation, and LC-MS quantification of malonyl-CoA. Western blotting and RT-qPCR confirm gene disruption and downstream effects. Functional assays for proliferation, migration, and drug response can be performed. The model also enables screening of metabolic drug targets and studies of lipid signaling in alveolar epithelium. For additional information, please contact Ascent Research.
