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Nat Biotechnol

Title:		An antisense-based functional genomics approach for identification of genes critical for growth of Candida albicans
Author(s):		De Backer MD; Nelissen B; Logghe M; Viaene J; Loonen I; Vandoninck S; de Hoogt R; Dewaele S; Simons FA; Verhasselt P; Vanhoof G; Contreras R; Luyten WH;
Address:		"Department of Advanced Bio-Technologies, Janssen Pharmaceutica, Turnhoutseweg 30, B2340 Beerse, Belgium. mdebacke@prius.jnj.com"
Journal Title:		Nat Biotechnol
Year:		2001
Volume:		19
Issue:		3
Page Number:		235 - 241
DOI:		10.1038/85677
ISSN/ISBN:		1087-0156 (Print) 1087-0156 (Linking)
Abstract:		"Converting the complete genome sequence of Candida albicans into meaningful biological information will require comprehensive screens for identifying functional classes of genes. Most systems described so far are not applicable to C. albicans because of its difficulty with mating, its diploid nature, and the lack of functional random insertional mutagenesis methods. We examined artificial gene suppression as a means to identify gene products critical for growth of this pathogen; these represent new antifungal drug targets. To achieve gene suppression we combined antisense RNA inhibition and promoter interference. After cloning antisense complementary DNA (cDNA) fragments under control of an inducible GAL1 promoter, we transferred the resulting libraries to C. albicans. Over 2,000 transformant colonies were screened for a promoter-induced diminished-growth phenotype. After recovery of the plasmids, sequence determination of their inserts revealed the messenger RNA (mRNA) they inhibited or the gene they disrupted. Eighty-six genes critical for growth were identified, 45 with unknown function. When used in high-throughput screening for antifungals, the crippled C. albicans strains generated in this study showed enhanced sensitivity to specific drugs"
Keywords:		"Antifungal Agents/pharmacology Candida albicans/drug effects/*genetics/*growth & development Cloning, Molecular/methods DNA, Antisense/genetics Drug Evaluation, Preclinical Fungal Proteins/genetics/metabolism Gene Expression Regulation, Fungal Gene Librar;"
Notes:		"MedlineDe Backer, M D Nelissen, B Logghe, M Viaene, J Loonen, I Vandoninck, S de Hoogt, R Dewaele, S Simons, F A Verhasselt, P Vanhoof, G Contreras, R Luyten, W H eng Research Support, Non-U.S. Gov't 2001/03/07 Nat Biotechnol. 2001 Mar; 19(3):235-41. doi: 10.1038/85677"