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Chapter 32 : Cool Tools 3: Large-Scale Genetic Interaction Screening in

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Cool Tools 3: Large-Scale Genetic Interaction Screening in , Page 1 of 2

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Abstract:

Although the genetic study of is generally less convenient than that of , recent advances in the molecular genetics of have greatly facilitated the study of this important human fungal pathogen. This chapter discusses the development of a novel strategy for large-scale synthetic genetic analysis in based on complex haploin-sufficiency (CHI). An actin null mutant () containing on a plasmid was mated to the set of single-gene-deletion strains and then cured of the plasmid to give a set of complex heterozygotes (). Many of the genes identified using this genetic approach demonstrated defects in the actin cytoskeleton, confirming the ability of CHI to identify interacting pairs of genes and genes with similar cellular functions. The development of a large-scale synthetic genetic screen using CHI to understand the role of the RAM network in morphological transition is described. As new mutagenesis techniques are developed and as large collections of mutants are created, it is likely that the creative application of these principles to genetic interaction analysis will lead to a deeper understanding of the complex regulatory networks that orchestrate biology and pathogenesis.

Citation: Chabrier-Roselló Y, Kumar A, Krysan D. 2012. Cool Tools 3: Large-Scale Genetic Interaction Screening in , p 497-500. In Calderone R, Clancy C (ed), and Candidiasis, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817176.ch32

Key Concept Ranking

Candida albicans
0.54285717
Saccharomyces cerevisiae
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0.54285717
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Image of FIGURE 1
FIGURE 1

Schematic depiction of simple haploinsufficiency and CHI. a* and b*, mutation in one allele of strain A or B in diploid . doi:10.1128/9781555817176.ch32.f1

Citation: Chabrier-Roselló Y, Kumar A, Krysan D. 2012. Cool Tools 3: Large-Scale Genetic Interaction Screening in , p 497-500. In Calderone R, Clancy C (ed), and Candidiasis, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817176.ch32
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Image of FIGURE 2
FIGURE 2

Schematic of screening strategy with in vitro mutagenesis of genomic library WO-1 using a Tn-based transposon containing the CaURA3-dpl200 auxotrophic marker. The resulting library was screened on Spider medium for altered filamentation relative to the parental strain. doi:10.1128/9781555817176.ch32.f2

Citation: Chabrier-Roselló Y, Kumar A, Krysan D. 2012. Cool Tools 3: Large-Scale Genetic Interaction Screening in , p 497-500. In Calderone R, Clancy C (ed), and Candidiasis, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817176.ch32
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Download as Powerpoint
Image of FIGURE 3
FIGURE 3

Venn diagram demonstrating the percentage of genes coregulated by RAM, PKA, or both pathways found through our CHI-based screen. doi:10.1128/9781555817176.ch32.f3

Citation: Chabrier-Roselló Y, Kumar A, Krysan D. 2012. Cool Tools 3: Large-Scale Genetic Interaction Screening in , p 497-500. In Calderone R, Clancy C (ed), and Candidiasis, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817176.ch32
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Download as Powerpoint

References

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