An improved tetracycline-inducible expression system for fission yeast

Abstract

AbstractThe ability to manipulate gene expression is valuable for elucidating gene function. In the fission yeastSchizosaccharomyces pombe, the most widely used regulatable expression system is thenmt1promoter and its two attenuated variants. However, these promoters have limitations, including a long lag, incompatibility with rich media, and unsuitability for non-dividing cells. Here, we present a tetracycline-inducible system free of these shortcomings. Our system features theenotetSpromoter, which achieves a similar induced level and a higher induction ratio compared to thenmt1promoter, without exhibiting a lag. Additionally, our system includes four weakenedenotetSvariants, offering an expression range similar to thenmt1series promoters but with more intermediate levels. To enhance usability, each promoter is combined with a Tet-repressor-expressing cassette in an integration plasmid. Importantly, our system can be used in non-dividing cells, enabling the development of a synchronous meiosis induction method with high spore viability. Moreover, our system allows for the shutdown of gene expression and the generation of conditional loss-of-function mutants. This system provides a versatile and powerful tool for manipulating gene expression in fission yeast.Summary statementA new inducible expression system for fission yeast enhances control, offers compatibility with non-dividing cells, and enables synchronous meiosis induction and conditional loss-of-function analysis.