Rapid colorimetric detection of genome evolution in SCRaMbLEd synthetic Saccharomyces Cerevisiae strains

Elizabeth L. I. Wightman, Heinrich Kroukamp*, Isak S. Pretorius, Ian T. Paulsen, Helena K. M. Nevalainen

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Abstract

Genome-scale engineering and custom synthetic genomes are reshaping the next generation of industrial yeast strains. The Cre-recombinase-mediated chromosomal rearrangement mechanism of designer synthetic Saccharomyces cerevisiae chromosomes, known as SCRaMbLE, is a powerful tool which allows rapid genome evolution upon command. This system is able to generate millions of novel genomes with potential valuable phenotypes, but the excessive loss of essential genes often results in poor growth or even the death of cells with useful phenotypes. In this study we expanded the versatility of SCRaMbLE to industrial strains, and evaluated different control measures to optimize genomic rearrangement, whilst limiting cell death. To achieve this, we have developed RED (rapid evolution detection), a simple colorimetric plate-assay procedure to rapidly quantify the degree of genomic rearrangements within a post-SCRaMbLE yeast population. RED-enabled semi-synthetic strains were mated with the haploid progeny of industrial yeast strains to produce stress-tolerant heterozygous diploid strains. Analysis of these heterozygous strains with the RED-assay, genome sequencing and custom bioinformatics scripts demonstrated a correlation between RED-assay frequencies and physical genomic rearrangements. Here we show that RED is a fast and effective method to evaluate the optimal SCRaMbLE induction times of different Cre-recombinase expression systems for the development of industrial strains.

Original languageEnglish
Article number1914
Pages (from-to)1-18
Number of pages18
JournalMicroorganisms
Volume8
Issue number12
DOIs
Publication statusPublished - 1 Dec 2020

Bibliographical note

Copyright the Author(s) 2020. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • Genome evolution
  • Industrial yeast strains
  • Saccharomyces cerevisiae
  • SCRaMbLE

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