The ACSF3 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited human cell population designed for functional studies of the ACSF3 gene. This product consists of a polyclonal pool of HeLa cells carrying diverse disruptions in the ACSF3 locus, enabling loss-of-function analysis without clonal artifacts. The heterogeneous knockout culture is suited for biochemical and metabolic profiling in a well-characterized cancerous epithelial background.
HeLa cells originate from human cervical adenocarcinoma and maintain HPV18 positivity; they are immortalized and extensively employed in biomedical research. Their robust proliferation and genetic tractability render them an ideal host for exploring mitochondrial fatty acid metabolism and disease-related pathway perturbations. The epithelial nature supports the study of metabolic disorders in a cellular context relevant to both cancer and inherited metabolic diseases.
ACSF3 encodes a mitochondrial malonyl-CoA synthetase that converts malonate to malonyl-CoA, providing the essential substrate for mitochondrial fatty acid synthesis (mtFAS) and influencing protein malonylation. The enzyme is regulated by upstream factors including PPARGC1A, SIRT1, and AMPK signaling, and it forms functional interactions with MCAT, NDUFAB1, and the mitochondrial acyl carrier protein. Within the mtFAS pathway, ACSF3 acts in concert with OXSM and MECR to generate fatty acyl chains, thereby impacting TCA cycle intermediate levels and post-translational modifications. Disruption of ACSF3 thus interrupts multiple facets of mitochondrial metabolism.
In the HeLa cell background, ACSF3 knockout provides a model to investigate combined malonic and methylmalonic aciduria (CMAMMA) and the broader role of mitochondrial malonyl-CoA metabolism in cancer biology. The absence of ACSF3 can perturb mitochondrial fatty acid synthesis, alter protein malonylation patterns, and shift energy homeostasis, making this model valuable for dissecting how cancer cells rely on mitochondrial anabolic pathways. This cellular context also facilitates exploration of potential therapeutic interventions targeting mitochondrial metabolism.
This product supports a range of assays, including LC-MS-based metabolic profiling to quantify malonate and malonyl-CoA pools, western blotting for ACSF3 and interacting partners like MCAT and NDUFAB1, RT-qPCR for transcriptional responses, and mitochondrial respiration assays using Seahorse analyzers. It also enables protein malonylation detection by immunoblotting or mass spectrometry and fatty acid synthesis assays with isotopic tracers, and is applicable to chemical screening and genetic rescue studies. For technical assistance, please contact Ascent Research.
