Bioorthogonal chemistry for deciphering lipid biology in living systems

Lipids are among the most structurally diverse classes of biomolecules and play fundamental roles in membrane organization, energy storage, and cellular signalling. However, unlike proteins and nucleic acids, they are not directly encoded by the genome, making their selective visualization, manipulation, and functional interrogation in living systems particularly challenging. Over the past two decades, bioorthogonal chemistry has transformed lipid research by enabling the incorporation of minimally perturbing chemical reporters and the use of highly selective ligation reactions in complex biological environments. These advances have made it possible to synthesize, label, image, quantify, and manipulate lipids with unprecedented spatial and temporal precision. In this review, we discuss how bioorthogonal chemistry has expanded the experimental toolkit for deciphering lipid biology, highlighting applications ranging from the in situ synthesis of membrane-forming lipids and live-cell imaging to the investigation of lipid-protein interactions and the development of bioinspired therapeutic strategies. Finally, we discuss the remaining challenges and emerging opportunities for achieving a quantitative, dynamic, and spatially resolved understanding of lipid function in living systems.