Carolacton, a macrocyclic natural product with impressive anti-biofilm biological activity, has been a focus in multiple research groups for the past decade. Chemists and biologists, alike, have been interested in uncovering the mechanism of action and have made great strides towards this goal. Carolacton causes cellular defects in Streptococcus mutans biofilm, which leads to decreases in cellular viability. As biological targets have been uncovered, synthetic chemists have devised synthetic routes that have helped uncover the important chemical functionalities that lead to biological activity. Herein, we discuss our synthetic collaboration that galvanized an entire research program around the natural product carolacton.