Competitive genomic screens of barcoded yeast libraries

Area of Science:

• Genomics
• Microbiology
• Functional Genomics

Background:

• Next-generation sequencing technologies provide daily access to new microbial genomes, necessitating faster methods for gene function determination.
• Traditional gene deletion mutant collections are not scalable for high-throughput analysis of thousands of genes.
• Existing parallelized assays rely on pre-existing barcoded mutant collections, limiting their application to well-characterized organisms.

Purpose of the Study:

• To bridge the gap between DNA sequencing and the creation of barcoded mutant collections for poorly characterized microbes.
• To develop a combined transposon disruption-barcoding approach for scalable functional genomics.
• To demonstrate the utility of this method for generating large-scale interaction datasets.

Main Methods:

• Developed a combined transposon disruption-barcoding strategy for introducing DNA tags into microbial genomes.
• Created a barcoded disruption collection for the fungal pathogen Candida albicans.
• Utilized both microarray and next-generation sequencing platforms for high-throughput barcode abundance analysis.

Main Results:

• Successfully generated a barcoded disruption collection for Candida albicans.
• Demonstrated the capability to collect 10,000 to 1,000,000 gene-gene and drug-gene interactions in a single experiment.
• Validated the scalability and efficiency of the transposon disruption-barcoding approach.

Conclusions:

• The combined transposon disruption-barcoding method significantly enhances the ability to perform parallel, high-throughput functional genomics assays in newly sequenced microbes.
• This approach facilitates rapid functional characterization and the discovery of gene-gene and drug-gene interactions.
• Enables the expansion of functional genomics studies to a wider range of microbial species.