covert genetic selections to optimize phenotypes隐性遗传选择优化表型.pdfVIP

Covert Genetic Selections to Optimize Phenotypes 1 2 2 Di Wu , Elizabeth Townsley , Alan Michael Tartakoff * 1 Monash Institute of Medical Research, Monash University, Monash Medical Centre, Melbourne, Victoria, Australia, 2 Department of Pathology, Cell Biology Program, Case Western Reserve University School of Medicine, Cleveland, Ohio, United States of America In many high complexity systems (cells, organisms, institutions, societies, economies, etc.), it is unclear which components should be regulated to affect overall performance. To identify and prioritize molecular targets which impact cellular phenotypes, we have developed a selection procedure (‘‘SPI’’–single promoting/inhibiting target identification) which monitors the abundance of ectopic cDNAs. We have used this approach to identify growth regulators. For this purpose, complex pools of S. cerevisiae cDNA transformants were established and we quantitated the evolution of the spectrum of cDNAs which was initially present. These data emphasized the importance of translation initiation and ER-Golgi traffic for growth. SPI provides functional insight into the stability of cellular phenotypes under circumstances in which established genetic approaches cannot be implemented. It provides a functional ‘‘synthetic genetic signature’’ for each state of the cell (i.e. genotype and environment) by surveying complex genetic libraries, and does not require specialized arrays of cDNAs/shRNAs, deletion strains, direct assessment of clonal growth or even a conditional phenotype. Moreover, it establishes a hierarchy of importance of those targets which can contribute, either positively or negatively, to modify the prevailing phenotype.