SATAY2.0

The analysis of large-scale transposon mutant libraries is becoming a method of choice for functional genomics in bacteria and fungi. We previously established SAturated Transposon Analysis in Yeast (SATAY), as a means to uncover the set of genes necessary for growth in any condition in S. cerevisiae. Here, we present an improved version of the protocol that leverages homologous recombination to increase transposition efficiency by a factor of 10. This improved protocol allows a single experimenter to perform several parallel screens in a short time span. We demonstrate its power by presenting (1) a comparison of the essential gene sets between two yeast laboratory backgrounds, (2) a comprehensive description of essential genes that display a phenotypic delay – we highlight the features that are common to these genes and propose plausible explanations for this phenomenon –, and (3) a genome-wide analysis of loss- and gain-of-function mutations conferring sensitivity or resistance to a compendium of 9 anti-fungal compounds. This study highlights the power of this improved SATAY protocol for yeast functional- and pharmaco-genomics.