Cytokinesis is the process by which cells physically separate after the duplication and spatial segregation of the genetic material. A number of general principles apply to this process. First the microtubule cytoskeleton plays an important role in the choice and positioning of the division site. Once the site is chosen, the local assembly of the actomyosin contractile ring remodels the plasma membrane. Finally, membrane trafficking to and membrane fusion at the division site cause the physical separation of the daughter cells, a process termed abscission. Here we will discuss recent advances in our understanding of the mechanisms of cytokinesis in animals, yeast, and plants.