High-throughput approaches to investigate neutral lipid biosynthesis

Rodrigo MP Siloto, Randall J Weselake

Agricultural Lipid Biotechnology Program, Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada

Abstract: Neutral lipids are ubiquitously found in nature, serving primarily as energy storage compounds. The ability to control the rate of neutral lipid bioassembly is of particular interest in numerous applications. For example, restriction of triacylglycerol and steryl ester synthesis in mammals could serve as adjuvant therapies against important diseases. In oilseeds, a boost in triacylglycerol accumulation would have a great impact in agriculture. Developing mechanisms to regulate neutral lipids synthesis requires rather specialized experimental designs where highthroughput screening approaches could be applied. Here we describe several enzyme targets that catalyze the bioassembly of neutral lipids, and potential applications for mechanisms that regulate their activity. We also describe different biochemical and cell-based assays to detect neutral lipid biosynthesis that are compatible with high-throughput formats. Several examples of established and prospective applications for these assays are discussed.

Keywords: neutral lipids, lipotoxicity, Nile red, scintillation proximity assay