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Abstract Steranes preserved in sedimentary rocks serve as molecular fossils, which are thought to record the expansion of eukaryote life through the Neoproterozoic Era ( ~ 1000-541 Ma). Scientists hypothesize that ancient C₂₇steranes originated from cholesterol, the major sterol produced by living red algae and animals. Similarly, C₂₈and C₂₉steranes are thought to be derived from the sterols of prehistoric fungi, green algae, and other microbial eukaryotes. However, recent work on annelid worms–an advanced group of eumetazoan animals–shows that they are also capable of producing C₂₈and C₂₉sterols. In this paper, we explore the evolutionary history of the24-C sterol methyltransferase(smt) gene in animals, which is required to make C₂₈₊sterols. We find evidence that thesmtgene was vertically inherited through animals, suggesting early eumetazoans were capable of C₂₈₊sterol synthesis. Our molecular clock of the animalsmtgene demonstrates that its diversification coincides with the rise of C₂₈and C₂₉steranes in the Neoproterozoic. This study supports the hypothesis that early eumetazoans were capable of making C₂₈₊sterols and that many animal lineages independently abandoned its biosynthesis around the end-Neoproterozoic, coinciding with the rise of abundant eukaryotic prey.