Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae

Publication Date:
2018-01-05
Publisher:
Genetics Society of America (GSA)
Electronic ISSN:
2160-1836
Topics:
Biology
Published by:
http://www.genetics-gsa.org/
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autor Xie, Z.-X., Mitchell, L. A., Liu, H.-M., Li, B.-Z., Liu, D., Agmon, N., Wu, Y., Li, X., Zhou, X., Li, B., Xiao, W.-H., Ding, M.-Z., Wang, Y., Yuan, Y.-J., Boeke, J. D.
beschreibung 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 HML α and HMR a 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.
citation_standardnr 6130017
datenlieferant ipn_articles
feed_id 169615
feed_publisher Genetics Society of America (GSA)
feed_publisher_url http://www.genetics-gsa.org/
insertion_date 2018-01-05
journaleissn 2160-1836
publikationsjahr_anzeige 2018
publikationsjahr_facette 2018
publikationsjahr_intervall 7984:2015-2019
publikationsjahr_sort 2018
publisher Genetics Society of America (GSA)
quelle G3: Genes, Genomes, Genetics
relation http://www.g3journal.org/cgi/content/short/8/1/173?rss=1
search_space articles
shingle_author_1 Xie, Z.-X., Mitchell, L. A., Liu, H.-M., Li, B.-Z., Liu, D., Agmon, N., Wu, Y., Li, X., Zhou, X., Li, B., Xiao, W.-H., Ding, M.-Z., Wang, Y., Yuan, Y.-J., Boeke, J. D.
shingle_author_2 Xie, Z.-X., Mitchell, L. A., Liu, H.-M., Li, B.-Z., Liu, D., Agmon, N., Wu, Y., Li, X., Zhou, X., Li, B., Xiao, W.-H., Ding, M.-Z., Wang, Y., Yuan, Y.-J., Boeke, J. D.
shingle_author_3 Xie, Z.-X., Mitchell, L. A., Liu, H.-M., Li, B.-Z., Liu, D., Agmon, N., Wu, Y., Li, X., Zhou, X., Li, B., Xiao, W.-H., Ding, M.-Z., Wang, Y., Yuan, Y.-J., Boeke, J. D.
shingle_author_4 Xie, Z.-X., Mitchell, L. A., Liu, H.-M., Li, B.-Z., Liu, D., Agmon, N., Wu, Y., Li, X., Zhou, X., Li, B., Xiao, W.-H., Ding, M.-Z., Wang, Y., Yuan, Y.-J., Boeke, J. D.
shingle_catch_all_1 Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
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 HML α and HMR a 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.
Xie, Z.-X., Mitchell, L. A., Liu, H.-M., Li, B.-Z., Liu, D., Agmon, N., Wu, Y., Li, X., Zhou, X., Li, B., Xiao, W.-H., Ding, M.-Z., Wang, Y., Yuan, Y.-J., Boeke, J. D.
Genetics Society of America (GSA)
2160-1836
21601836
shingle_catch_all_2 Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
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 HML α and HMR a 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.
Xie, Z.-X., Mitchell, L. A., Liu, H.-M., Li, B.-Z., Liu, D., Agmon, N., Wu, Y., Li, X., Zhou, X., Li, B., Xiao, W.-H., Ding, M.-Z., Wang, Y., Yuan, Y.-J., Boeke, J. D.
Genetics Society of America (GSA)
2160-1836
21601836
shingle_catch_all_3 Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
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 HML α and HMR a 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.
Xie, Z.-X., Mitchell, L. A., Liu, H.-M., Li, B.-Z., Liu, D., Agmon, N., Wu, Y., Li, X., Zhou, X., Li, B., Xiao, W.-H., Ding, M.-Z., Wang, Y., Yuan, Y.-J., Boeke, J. D.
Genetics Society of America (GSA)
2160-1836
21601836
shingle_catch_all_4 Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
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 HML α and HMR a 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.
Xie, Z.-X., Mitchell, L. A., Liu, H.-M., Li, B.-Z., Liu, D., Agmon, N., Wu, Y., Li, X., Zhou, X., Li, B., Xiao, W.-H., Ding, M.-Z., Wang, Y., Yuan, Y.-J., Boeke, J. D.
Genetics Society of America (GSA)
2160-1836
21601836
shingle_title_1 Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
shingle_title_2 Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
shingle_title_3 Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
shingle_title_4 Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
timestamp 2025-06-30T23:31:46.200Z
titel Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
titel_suche Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of Saccharomyces cerevisiae
topic W
uid ipn_articles_6130017