Douglas AC, et al. (2012) Functional analysis with a barcoder yeast gene overexpression system. G3 (Bethesda) 2(10):1279-89
Abstract: 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.
|Status: Published||Type: Journal Article||PubMed ID: 23050238|
Topics addressed in this paper
Number of different genes curated to this paper: 15
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|Topics||Topics not linked to Genes||Genes (#1 - 10 )|
|Large-scale genetic interaction|
|Large-scale phenotype analysis|
|Techniques and Reagents|
|Topics||Genes (#11 - 15 )|