The ACACA Knockout HeLa Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the ACACA gene in the HeLa host background. This heterogeneous pool of edited cells is designed for loss-of-function investigations into acetyl-CoA carboxylase alpha (ACC??) biology, enabling researchers to interrogate the functional consequences of ACACA ablation without selecting clonal isolates. The polyclonal format captures a diverse spectrum of editing events, facilitating robust and reproducible examination of ACC??-dependent cellular processes.
The host cell line, HeLa, is a widely utilized human cervical adenocarcinoma model originally derived from an HPV18-positive tumor. These epithelial cells exhibit rapid proliferation and have been extensively characterized in cancer biology, virology, and metabolic research. HeLa cells are known to maintain active lipogenic programs, making them a pertinent system for exploring the role of de novo fatty acid synthesis in transformed cells. Their well-documented genetic and phenotypic traits offer a standardized platform for knockout studies.
ACACA encodes ACC??, the rate-limiting enzyme that catalyzes the ATP-dependent carboxylation of acetyl-CoA to malonyl-CoA, the critical committed step in de novo lipogenesis. ACC?? activity is tightly regulated: it is inhibited by AMPK-mediated phosphorylation and activated by insulin-stimulated dephosphorylation via phosphatase PP2A. Transcriptionally, SREBP1 upregulates ACACA expression, while citrate serves as an allosteric activator. Downstream, malonyl-CoA serves as a substrate for fatty acid synthase (FASN) and concurrently inhibits carnitine palmitoyltransferase 1 (CPT1), thereby modulating fatty acid oxidation. ACC?? interacts with biotin as a necessary cofactor and with ACACB, the minor isoform, within the broader acetyl-CoA metabolic network.
In the HeLa cellular context, ACACA disruption profoundly impacts lipid metabolism and may unmask dependencies on exogenous lipids or alternative metabolic pathways. Given that many cancers, including cervical carcinoma, upregulate lipogenesis to support membrane biosynthesis and proliferation, this knockout model provides a tractable system to dissect how loss of ACC?? activity alters cancer cell fitness. It also permits investigation of the interplay between HPV18 oncoproteins and host cell metabolism, as HPV-driven cells often rewire metabolic circuits.
Researchers can employ this ACACA knockout polyclonal population in diverse applications, including cancer metabolism studies, lipogenesis inhibition screens, AMPK signaling analyses, and metabolic disease modeling. Representative assays compatible with this model include Western blotting to confirm loss of ACACA and downstream FASN expression, [14C]-acetate incorporation to measure de novo fatty acid synthesis, lipid droplet staining with neutral lipid dyes, Seahorse metabolic flux analysis to profile oxidative and glycolytic metabolism, and flow cytometry for quantitative lipid content. For additional details or to explore how this reagent can advance your research, please contact Ascent Research.
