The Saccharomyces cerevisiae sequencing project (the first eukaryotic genome decoded) was completed in 1995 and, subsequently, the first version of the yeast knockout collection was made available in 2002. Since then, many diverse studies have applied these resources to understand drug mechanism of action and to identify novel drug targets and target pathways. In this update of an earlier review, we present a snapshot of the current state of chemical genomic approaches in yeast, propose a set of integrated chemical genomic assays to move the field forward and consider its near-term future.