A strategy has been developed for creating Saccharomyces cerevisiae strains with a high RNA content by following a three-step breeding procedure. In the first step, an S. cerevisiae disruptant of the RRN10 gene, one of the components of the UAF (upstream activation factor) complex of rRNA transcription, was constructed and showed severely slow growth. In the second step, seven suppressors were isolated that restored the slow growth of the Delta rrn10 disruptant Genetic analysis revealed that each of the seven suppressors that were isolated appeared to have dominant and multiple mutations. The specific growth rate of those suppressors was increased approximately two-fold as compared with the Delta rrn10 parental strain. The absolute RNA content showed that the suppressors had an RNA content 32-56% higher than that of the Delta rrn10 parental strain. In the last step, the RRN10 wild-type gene was integrated into chromosome V of each of the original suppressors. The total RNA content of the integrants was also 1.4- to 2.3-fold higher than the wild-type strain. In conclusion, since yeast RNA is the source of 5'-IMP and 5'-GMP that enhance the delicious taste in certain types of food, like soups and sauces, the strategy taken in this study provides effective approach to breed S. cerevisiae strains producing a higher content of RNA that will contribute to yeast food biotechnology. (C) 2011, The Society for Biotechnology, Japan. All rights reserved.