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Medientyp:
E-Artikel
Titel:
Functional Analysis With a Barcoder Yeast Gene Overexpression System
Beteiligte:
Douglas, Alison C;
Smith, Andrew M;
Sharifpoor, Sara;
Yan, Zhun;
Durbic, Tanja;
Heisler, Lawrence E;
Lee, Anna Y;
Ryan, Owen;
Göttert, Hendrikje;
Surendra, Anu;
van Dyk, Dewald;
Giaever, Guri;
Boone, Charles;
Nislow, Corey;
Andrews, Brenda J
Erschienen:
Oxford University Press (OUP), 2012
Erschienen in:G3 Genes|Genomes|Genetics
Sprache:
Englisch
DOI:
10.1534/g3.112.003400
ISSN:
2160-1836
Entstehung:
Anmerkungen:
Beschreibung:
<jats:title>Abstract</jats:title>
<jats:p>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.</jats:p>