RNA functional profiling by gene deletion in a model organism. (360G-Wellcome-094225_Z_10_Z)

Saccharomyces cerevisiae is a key model organism for investigating biological processes. The Yeast Deletion Collection (YDC) for protein-coding genes has provided a valuable resource for researchers. While it is important to determine the role protein-coding genes play in biological processes, RNA is increasingly playing a central role in many biological processes. Unfortunately, RNA-coding genes are absent from the YDC. Therefore, it is essential to develop deletion strains of RNA-coding genes to provide a comprehensive gene deletion collection for genome-scale analysis and systematic analysis of RNA function. We propose to delete all annotated and a collection of recently identified unannotated RNA genes in yeast. High-throughput deletion will include unique molecular barcodes allowing identification of each deletion strain, parallel analysis and YDC compatibility. The RNA deletion strains will be used for RNA functional analysis and to probe, by next generation sequencing of barcod es, the contribution of RNA to growth of yeast in continuous culture with a variety of conditions and treatments. Selected essential RNA genes will also be deleted in the divergent yeast Sigma1278b to determine whether phenotype is influenced by genetic background. Finally, strains will be distributed to international repositories to provide a lasting and useful resource for researchers.