Yeast Genetics to Investigate the Function of Core Pre-mRNA Splicing Factors

The yeast Saccharomyces cerevisiae has been used extensively to investigate the functions of RNA and protein components of the spliceosome. As many of the yeast core splicing factors are evolutionarily conserved with human counterparts, information gained in yeast can be applied directly to humans. The ease with which budding yeast can be manipulated genetically has resulted in numerous resources that make genetic analysis in this organism routine. The function of any gene that codes for a spliceosomal component can be analyzed in yeast, and its genetic interactions investigated using genetic screens. Synthetic enhancement or suppression occur when a mutation in one gene respectively exacerbates or alleviates the severity of a mutation in another gene, indicating a functional interaction. In the case of splicing factors, this can lead to new information on the mechanisms of splicing. In this chapter, methods are described for the functional analysis and synthetic enhancement/lethality screens in yeast. © 2012 Wiley-VCH Verlag GmbH & Co. KGaA.