Abstract
A valuable approach to understanding small non-coding RNA (sncRNA) function is to regulate their expression. However, inducible expression of sncRNAs can be challenging. Here, we present a protocol to express sncRNAs in Saccharomyces cerevisiae using a two-plasmid inducible tet-on system. We describe the steps for generating plasmids and inducing and assessing sncRNA expression. This protocol can be used to express and elucidate the function of tRNA introns and has the potential to identify functions of a wide range of sncRNAs. For complete details on the use and execution of this protocol, please refer to Nostramo et al.(1).