Expanding room for tetrazine ligations in the in vivo chemistry toolbox

There is tremendous interest in developing and refining methods to predictably perform chemical reactions within the framework of living systems. Here we review recent advances in applying tetrazine cycloadditions to live cell and in vivo chemistry. We highlight new syntheses of the tetrazine and dienophile precursors useful for in vivo studies. We briefly overview the use of this reaction in combination with unnatural amino acid technology and discuss applications involving the imaging of glycans on live cells. An emerging area is the use of tetrazine ligations for the development of in vivo imaging probes such as those used for positron emission tomography. We summarize recent applications involving tetrazine cycloadditions performed in live mice for pretargeted imaging of cancer cell biomarkers.