Functional Genomics by Transposon Tagging in Yeast

Anuj Kumar
Project Manager, Yale Genome Analysis Center
Department of Molecular, Cellular and Developmental Biology
Yale University
P. O. Box 208103
New Haven, CT 06520-8103
telephone: 203-429-9949
fax: 203-432-6161
prestype: Platform
presenter: Anuj Kumar

Transposable elements are powerful tools for insertional mutagenesis, notable for the scale of throughput and ease of analysis they offer. As a result, transposon-based systems provide an excellent method of genome-wide mutagenesis. Illustrating this point, we have employed transposon mutagenesis as a means of elucidating gene function on a genome-wide scale in Saccharomyces cerevisiae. Our approach is unique in its versatility: by designing specially modified multi-functional mini-transposons, we can measure gene expression, generate gene disruptions, and localize gene products; all from a single transposon insertion event. Using these multi-functional transposons, we have generated a collection of roughly 17,000 yeast mutants, each carrying a single transposon insertion within a region of the genome expressed during vegetative growth and/or sporulation. This collection has been used to determine disruption phenotypes for nearly 8,000 strains under 20 different growth conditions. Additionally, we have identified over 300 previously non-annotated open reading frames and determined the subcellular localization of transposon-tagged proteins in approximately 9,000 yeast mutants. In total, our study encompasses over a quarter of a million data points, providing insight into the function of nearly 3,000 yeast genes. In addition to the wealth of data generated by this approach, our genome-wide collections of defined mutants constitute a valuable laboratory reagent, streamlining future genetic screens by eliminating the need to identify the affected gene within a given mutant of interest. As such, these collections of defined alleles promise to greatly expedite studies of gene function on a genomic scale.

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