Functional Analysis With a Barcoder Yeast Gene Overexpression System

G3: Genes, Genomes, Genetics

Guri Giaever¹  Andrew M. Smith⁵  Hendrikje Göttert⁵  Dewald van Dyk³  Brenda J. Andrews⁵  Corey Nislow⁵  Lawrence E. Heisler²  Anu Surendra⁴  Sara Sharifpoor⁴  Charles Boone⁵  Anna Y. Lee²  Alison C. Douglas⁵  Zhun Yan²  Tanja Durbic⁴  Owen Ryan⁵ 

[1] Department of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Pharmaceutical Sciences, University of Toronto, Ontario M5S 3M2, CDepartment of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CDepartment of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Pharmaceutical Sciences, University of Toronto, Ontario M5S 3M2, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Pharmaceutical Sciences, University of Toronto, Ontario M5S 3M2, CDepartment of Pharmaceutical Sciences, University of Toronto, Ontario M5S 3M2, CDepartment of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Pharmaceutical Sciences, University of Toronto, Ontario M5S 3M2, C;Donnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Pharmaceutical Sciences, University of Toronto, Ontario M5S 3M2, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Pharmaceutical Sciences, University of Toronto, Ontario M5S 3M2, CDepartment of Pharmaceutical Sciences, University of Toronto, Ontario M5S 3M2, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Pharmaceutical Sciences, University of Toronto, Ontario M5S 3M2, C;Banting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CBanting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, CBanting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CBanting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, C;Donnelly Centre, University of Toronto, Ontario M5S 3E1, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, C;Department of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CBanting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CDepartment of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CBanting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CBanting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, CDepartment of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CBanting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, CDepartment of Molecular Genetics, University of Toronto, Ontario M5S 1A8, CBanting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, CDonnelly Centre, University of Toronto, Ontario M5S 3E1, C

关键词: barFLEX array;    gene overexpression;    barcoders;    synthetic dosage lethality;   

DOI  :  10.1534/g3.112.003400

学科分类：生物科学（综合）

来源: Genetics Society of America

【 摘 要 】

Systematic analysis of gene overexpression phenotypes provides an insight into gene function, enzyme targets, and biological pathways. Here, we describe a novel functional genomics platform that enables a highly parallel and systematic assessment of overexpression phenotypes in pooled cultures. First, we constructed a genome-level collection of ~5100 yeast barcoder strains, each of which carries a unique barcode, enabling pooled fitness assays with a barcode microarray or sequencing readout. Second, we constructed a yeast open reading frame (ORF) galactose-induced overexpression array by generating a genome-wide set of yeast transformants, each of which carries an individual plasmid-born and sequence-verified ORF derived from the Saccharomyces cerevisiae full-length EXpression-ready (FLEX) collection. We combined these collections genetically using synthetic genetic array methodology, generating ~5100 strains, each of which is barcoded and overexpresses a specific ORF, a set we termed “barFLEX.” Additional synthetic genetic array allows the barFLEX collection to be moved into different genetic backgrounds. As a proof-of-principle, we describe the properties of the barFLEX overexpression collection and its application in synthetic dosage lethality studies under different environmental conditions.

【 授权许可】

Unknown