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http://dx.doi.org/10.4014/jmb.0806.398

Quantifiable Downregulation of Endogenous Genes in Agaricus bisporus Mediated by Expression of RNA Hairpins  

Costa, Ana S.M.B. (Warwick HRI, University of Warwick)
Thomas, D. John I. (Warwick HRI, University of Warwick)
Eastwood, Daniel (Warwick HRI, University of Warwick)
Cutler, Simon B. (Warwick HRI, University of Warwick)
Bailey, Andy M. (School of Biological Sciences, University of Bristol)
Foster, Gary D. (School of Biological Sciences, University of Bristol)
Mills, Peter R. (Warwick HRI, University of Warwick)
Challen, Michael P. (Warwick HRI, University of Warwick)
Publication Information
Journal of Microbiology and Biotechnology / v.19, no.3, 2009 , pp. 271-276 More about this Journal
Abstract
Functional gene studies in the cultivated white button mushroom Agaricus bisporus have been constrained by the absence of effective gene-silencing tools. Using two endogenous genes from A. bisporus, we have tested the utility of dsRNA hairpin constructs to mediate downregulation of specific genes. Hairpin constructs for genes encoding orotidine 5'-monophosphate decarboxylase (URA3) and carboxin resistance (CBX) were introduced into A. bisporus using Agrobacteriummediated transfection. Although predicted changes in phenotype were not observed in vitro, quantitative-PCR analyses indicated unambiguously that transcripts in several transformants were substantially reduced compared with the non-transformed controls. Interestingly, some hairpin transformants exhibited increased transcription of target genes. Our observations show that hairpin transgenic sequences can mediate downregulation of A. bisporus endogenous genes and that the technology has the potential to expedite functional genomics of the mushroom.
Keywords
Agaricus bisporus; CBX; gene-silencing; dsRNA hairpin; RNAi; URA3;
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