Systematic Complex Haploinsufficiency-Based Genetic Analysis of Candida albicans Transcription Factors: Tools and Applications to Virulence-Associated Phenotypes

G3 : Genes - Genomes - Genetics
Virginia E GlazierDamian J Krysan

Abstract

Genetic interaction analysis is a powerful approach to the study of complex biological processes that are dependent on multiple genes. Because of the largely diploid nature of the human fungal pathogen Candida albicans, genetic interaction analysis has been limited to a small number of large-scale screens and a handful for gene-by-gene studies. Complex haploinsufficiency, which occurs when a strain containing two heterozygous mutations at distinct loci shows a phenotype that is distinct from either of the corresponding single heterozygous mutants, is an expedient approach to genetic interactions analysis in diploid organisms. Here, we describe the construction of a barcoded-library of 133 heterozygous TF deletion mutants and deletion cassettes for designed to facilitate complex haploinsufficiency-based genetic interaction studies of the TF networks in C. albicans We have characterized the phenotypes of these heterozygous mutants under a broad range of in vitro conditions using both agar-plate and pooled signature tag-based assays. Consistent with previous studies, haploinsufficiency is relative uncommon. In contrast, a set of 12 TFs enriched in mutants with a role in adhesion were found to have altered competitive fitness at ea...Continue Reading

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Citations

Mar 19, 2019·Proteomics·Mark R BleackleyMarilyn A Anderson
Dec 13, 2019·PLoS Biology·Alicia Sanchez-GorostiagaAlvaro Sanchez
Jan 17, 2020·PLoS Pathogens·Florent MorioGeraldine Butler
Apr 24, 2020·PLoS Pathogens·Virginia E Glazier, Damian J Krysan
Oct 31, 2018·Nature Communications·Jiaxin GaoJianbin Wang
Jan 2, 2020·Journal of Fungi·Shen-Huan Liang, Richard J Bennett
Jan 26, 2021·Current Protocols·Manning Y HuangAaron P Mitchell
Oct 31, 2021·Nature Communications·Robert ZarnowskiDavid R Andes
Dec 6, 2021·FEMS Yeast Research·Nicholas C GervaisRebecca S Shapiro

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Methods Mentioned

BETA
PCR
light microscopy

Software Mentioned

ImageStream
Elements
NIS
IDEAS
INSPIRE

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