Genome-wide construction of a series of designed segmental aneuploids in Saccharomyces cerevisiae

Abstract

Segmental aneuploidy can play an important role in environmental adaptation. However, study of segmental aneuploids is severely hampered by the difficulty of creating them in a designed fashion. Here, we describe a PCR-mediated chromosome duplication (PCDup) technology that enables the generation of segmental aneuploidy at any desired chromosomal region in Saccharomyces cerevisiae. We constructed multiple strains harboring 100 kb to 200 kb segmental duplications covering the whole of the S. cerevisiae genome. Interestingly, some segmental aneuploidies confer stress tolerance, such as to high temperature, ethanol and strong acids, while others induce cell lethality and stress sensitivity, presumably as result of the simultaneous increases in dosages of multiple genes. We suggest that our PCDup technology will accelerate studies into the phenotypic changes resulting from alteration of gene dosage balance of multiple genes and will provide new insights into the adaptive molecular mechanisms in the genome in segmental aneuploidy-derived human diseases.

Systematic approach for assessing whether undeletable chromosomal regions in Saccharomyces cerevisiae are required for cell viability

“…Splitting modules were prepared according to Sasano et al (2016) by using pSJ70, pSJ23 and p3121 as template plasmids. The duplication module was prepared according to Natesuntorn et al (2015) with slight modification. Specifically, rather than a 400 bp homology region used by Natesuntorn et al (2015), we used a 50 bp homology sequence to duplicate the target regions.…”

“…The duplication module was prepared according to Natesuntorn et al (2015) with slight modification. Specifically, rather than a 400 bp homology region used by Natesuntorn et al (2015), we used a 50 bp homology sequence to duplicate the target regions. Primers used for making DNA modules for replacement, splitting or duplicating target regions are listed in Additional file 1: Table S1.…”

Yeasts for Global Happiness: report of the 14th International Congress on Yeasts (ICY14) held in Awaji Island

“…; Natesuntorn et al . ) (day 1); (ii) ‘A multi‐faceted cellular response against aneuploidy stress’ by Dr. Davis Ng (National University of Singapore, Singapore; Fig. D) regarding biochemical and genomewide analyses of cellular responses induced by genomic imbalance from the perspective of proteotoxic stress (Thibault & Ng ) (day 2); (iii) ‘Yeast as a model organism in ecology and evolution’ by Dr. Graham Bell (McGill University, Canada; Fig.…”

“…The plenary lectures of ICY14 consisted of: (i) 'Yeast genome engineering -A new challenge to deciphering genome function and breeding' by Dr. Satoshi Harashima (Sojo University, Japan; Fig. 1C) regarding the frontier of genome engineering technologies for bioscience and biotechnology (Sugiyama et al 2009;Natesuntorn et al 2015) (day 1); (ii) 'A multifaceted cellular response against aneuploidy stress' by Dr. Davis Ng (National University of Singapore, Singapore; Fig. 1D) regarding biochemical and genomewide analyses of cellular responses induced by genomic imbalance from the perspective of proteotoxic stress (Thibault & Ng 2012) (day 2); (iii) 'Yeast as a model organism in ecology and evolution' by Dr. Graham Bell (McGill University, Fig.…”

CRISPR-PCDup: a novel approach for simultaneous segmental chromosomal duplication in Saccharomyces cerevisiae

“…Previously, we developed a variety of new chromosome engineering technologies in S. cerevisiae. One such method, named PCR-mediated chromosome duplication (PCDup), enables the duplication of any desired chromosomal region as an independent chromosome (Natesuntorn et al 2015). PCDup is able to duplicate chromosomal regions with lengths from 50 kb to 300 kb.…”