@article{0f9843a6a4ab402d8b91580cd37cb4d9,
title = "Rapid and efficient CRISPR/Cas9-based mating-type switching of Saccharomyces cerevisiae",
abstract = "Rapid and highly efficient mating-type switching of Saccharomyces cerevisiae enables a wide variety of genetic manipulations, such as the construction of strains, for instance, isogenic haploid pairs of both mating-types, diploids and polyploids. We used the CRISPR/Cas9 system to generate a double-strand break at the MAT locus and, in a single cotransformation, both haploid and diploid cells were switched to the specified mating-type at ~80% efficiency. The mating-type of strains carrying either rod or ring chromosome III were switched, including those lacking HMLa and HMRa cryptic mating loci. Furthermore, we transplanted the synthetic yeast chromosome V to build a haploid polysynthetic chromosome strain by using this method together with an endoreduplication intercross strategy. The CRISPR/Cas9 mating-type switching method will be useful in building the complete synthetic yeast (Sc2.0) genome. Importantly, it is a generally useful method to build polyploids of a defined genotype and generally expedites strain construction, for example, in the construction of fully a/a/α/α isogenic tetraploids.",
keywords = "CRISPR/Cas9, Mating-type switching, Polyploidy, Ring chromosome, Saccharomyces cerevisiae",
author = "Xie, {Ze Xiong} and Mitchell, {Leslie A.} and Liu, {Hui Min} and Li, {Bing Zhi} and Duo Liu and Neta Agmon and Yi Wu and Xia Li and Xiao Zhou and Bo Li and Xiao, {Wen Hai} and Ding, {Ming Zhu} and Ying Wang and Yuan, {Ying Jin} and Boeke, {Jef D.}",
note = "Funding Information: to the Sc2.0 consortium. We dedicate this paper to the memory of Amar Klar. The work in China was supported by the Ministry of Science and Technology of China (grants 2014CB745100 and 2015DFA00960), and the National Natural Science Foundation of China (grants 21390203 and 21621004). The work in the United States was supported by National Science Foundation grants MCB-1026068 and MCB-1158201 to J.D.B. Funding Information: We thank Rodney Rothstein for the generous contribution of the set of centromere destabilizing constructs, originally developed in his laboratoryto the Sc2.0 consortium. We dedicate this paper to the memory of Amar Klar. The work in China was supported by the Ministry of Science and Technology of China (grants 2014CB745100 and 2015DFA00960), and the National Natural Science Foundation of China (grants 21390203 and 21621004). The work in the United States was supported by National Science Foundation grants MCB-1026068 and MCB-1158201 to J.D.B. Publisher Copyright: {\textcopyright} 2018 Xie et al.",
year = "2018",
month = jan,
day = "1",
doi = "10.1534/g3.117.300347",
language = "English (US)",
volume = "8",
pages = "173--183",
journal = "G3 (Bethesda, Md.)",
issn = "2160-1836",
publisher = "Genetics Society of America",
number = "1",
}