Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic <i>Saccharomyces cerevisiae</i> Strains
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 <i>Saccharomyces cerevisiae</i> chromosomes, known as SCRaMbLE, is a powerful tool w...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-12-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/8/12/1914 |
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| author | Elizabeth L. I. Wightman Heinrich Kroukamp Isak S. Pretorius Ian T. Paulsen Helena K. M. Nevalainen |
| author_facet | Elizabeth L. I. Wightman Heinrich Kroukamp Isak S. Pretorius Ian T. Paulsen Helena K. M. Nevalainen |
| author_sort | Elizabeth L. I. Wightman |
| collection | DOAJ |
| description | 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 <i>Saccharomyces cerevisiae</i> 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. |
| first_indexed | 2024-03-10T14:24:36Z |
| format | Article |
| id | doaj.art-b8faf730f7554bddaa56efe30b7d8376 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T14:24:36Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-b8faf730f7554bddaa56efe30b7d83762023-11-20T23:08:12ZengMDPI AGMicroorganisms2076-26072020-12-01812191410.3390/microorganisms8121914Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic <i>Saccharomyces cerevisiae</i> StrainsElizabeth L. I. Wightman0Heinrich Kroukamp1Isak S. Pretorius2Ian T. Paulsen3Helena K. M. Nevalainen4Centre of Excellence in Synthetic Biology, Department of Molecular Sciences, Macquarie University, Sydney, NSW 2109, AustraliaCentre of Excellence in Synthetic Biology, Department of Molecular Sciences, Macquarie University, Sydney, NSW 2109, AustraliaChancellery, Macquarie University, Sydney, NSW 2109, AustraliaCentre of Excellence in Synthetic Biology, Department of Molecular Sciences, Macquarie University, Sydney, NSW 2109, AustraliaCentre of Excellence in Synthetic Biology, Department of Molecular Sciences, Macquarie University, Sydney, NSW 2109, AustraliaGenome-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 <i>Saccharomyces cerevisiae</i> 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.https://www.mdpi.com/2076-2607/8/12/1914<i>Saccharomyces cerevisiae</i>SCRaMbLEgenome evolutionindustrial yeast strains |
| spellingShingle | Elizabeth L. I. Wightman Heinrich Kroukamp Isak S. Pretorius Ian T. Paulsen Helena K. M. Nevalainen Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic <i>Saccharomyces cerevisiae</i> Strains Microorganisms <i>Saccharomyces cerevisiae</i> SCRaMbLE genome evolution industrial yeast strains |
| title | Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic <i>Saccharomyces cerevisiae</i> Strains |
| title_full | Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic <i>Saccharomyces cerevisiae</i> Strains |
| title_fullStr | Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic <i>Saccharomyces cerevisiae</i> Strains |
| title_full_unstemmed | Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic <i>Saccharomyces cerevisiae</i> Strains |
| title_short | Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic <i>Saccharomyces cerevisiae</i> Strains |
| title_sort | rapid colorimetric detection of genome evolution in scrambled synthetic i saccharomyces cerevisiae i strains |
| topic | <i>Saccharomyces cerevisiae</i> SCRaMbLE genome evolution industrial yeast strains |
| url | https://www.mdpi.com/2076-2607/8/12/1914 |
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