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The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions

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Published on: December 15, 2012

An Optimized Genotyping Workflow for Identifying Highly SCRaMbLEd Synthetic Yeasts.

Timon A Lindeboom¹, María Del Carmen Sanchez Olmos¹, Karina Schulz²

¹Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Str. 10, 35043 Marburg, Germany.

ACS Synthetic Biology

|April 10, 2024

Summary

This summary is machine-generated.

Synthetic yeast strains (Sc2.0) can generate genomic diversity using SCRaMbLE technology. New qPCR primers (loxTags) and a primer prediction tool (qTagGer) enable efficient genotyping of these diverse yeast populations.

Keywords:

PCRTags SCRaMbLE genome rearrangements optogenetics qPCR genotyping recombinases synthetic biology synthetic yeast

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Last Updated: Jun 28, 2025

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Published on: March 7, 2019

Area of Science:

Synthetic biology
Genomics
Molecular biology

Background:

Synthetic yeast strains (Sc2.0) possess numerous recombination sites (loxPsym) enabling genome restructuring via SCRaMbLE.
SCRaMbLE technology can generate highly diverse yeast populations from a single genotype.

Purpose of the Study:

To develop an efficient workflow for selecting genetically diverse yeast candidates with rearranged synthetic chromosomes.
To introduce loxTags, qPCR primers for genotyping loxPsym sites, and qTagGer, a primer prediction tool.

Main Methods:

Developed loxTags, a set of qPCR primers for genotyping across loxPsym sites.
Created qTagGer, a qPCR genotyping primer prediction tool to manage numerous amplicons.
Utilized loxTag-based genotyping and long-read sequencing.

Main Results:

loxTags can detect deletions, inversions, and translocations after SCRaMbLE.
Light-inducible Cre recombinase (L-SCRaMbLE) efficiently generated diverse recombination events in Sc2.0 strains.
Demonstrated successful application on synthetic chromosome III (linear and circular versions).

Conclusions:

loxTags and qTagGer provide an efficient workflow for analyzing SCRaMbLE-induced genomic diversity in synthetic yeast.
L-SCRaMbLE is effective in generating complex genomic rearrangements in Sc2.0 yeast strains.