CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae
In this study, we focused on the applicability of CRISPR/Cpf1 in genome simplification of Saccharomyces cerevisiae and established a CRISPR/Cpf1 assisted method for rapid markerless large fragment deletion to facilitate laboratory evolution of geome of S. cerevisiae by rational genetic engineering....
| Main Authors: | , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
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2020
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| Online Access: | https://hdl.handle.net/10356/139529 |
| _version_ | 1826116773397659648 |
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| author | Li, Zhen-Hai Liu, Min Lyu, Xiao-Mei Wang, Feng-Qing Wei, Dong-Zhi |
| author2 | School of Chemical and Biomedical Engineering |
| author_facet | School of Chemical and Biomedical Engineering Li, Zhen-Hai Liu, Min Lyu, Xiao-Mei Wang, Feng-Qing Wei, Dong-Zhi |
| author_sort | Li, Zhen-Hai |
| collection | NTU |
| description | In this study, we focused on the applicability of CRISPR/Cpf1 in genome simplification of Saccharomyces cerevisiae and established a CRISPR/Cpf1 assisted method for rapid markerless large fragment deletion to facilitate laboratory evolution of geome of S. cerevisiae by rational genetic engineering. This method uses a Cpf1 expression plasmid and a crRNA array expression plasmid. The DNA fragment between two DSBs generated by CRISPR/Cpf1 can be cut off from the chromosome, along with re-ligation of the genomic endpoints of the DSBs. Using this method, the large DNA fragment of ∼38 kb between the two genes of TRM10 and REX4 was successfully and rapidly deleted, which was verified by PCR and Sanger DNA Sequencing. This method is simple and rapid, and can be easily implemented for large fragment deletion in S. cerevisiae. |
| first_indexed | 2024-10-01T04:17:15Z |
| format | Journal Article |
| id | ntu-10356/139529 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:17:15Z |
| publishDate | 2020 |
| record_format | dspace |
| spelling | ntu-10356/1395292020-05-20T04:00:22Z CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae Li, Zhen-Hai Liu, Min Lyu, Xiao-Mei Wang, Feng-Qing Wei, Dong-Zhi School of Chemical and Biomedical Engineering Engineering::Bioengineering CRISPR/Cpf1 Large Fragment Deletion In this study, we focused on the applicability of CRISPR/Cpf1 in genome simplification of Saccharomyces cerevisiae and established a CRISPR/Cpf1 assisted method for rapid markerless large fragment deletion to facilitate laboratory evolution of geome of S. cerevisiae by rational genetic engineering. This method uses a Cpf1 expression plasmid and a crRNA array expression plasmid. The DNA fragment between two DSBs generated by CRISPR/Cpf1 can be cut off from the chromosome, along with re-ligation of the genomic endpoints of the DSBs. Using this method, the large DNA fragment of ∼38 kb between the two genes of TRM10 and REX4 was successfully and rapidly deleted, which was verified by PCR and Sanger DNA Sequencing. This method is simple and rapid, and can be easily implemented for large fragment deletion in S. cerevisiae. 2020-05-20T04:00:22Z 2020-05-20T04:00:22Z 2018 Journal Article Li, Z.-H., Liu, M., Lyu, X.-M., Wang, F.-Q., & Wei, D.-Z. (2018). CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae. Journal of Basic Microbiology, 58(12), 1100-1104. doi:10.1002/jobm.201800195 0233-111X https://hdl.handle.net/10356/139529 10.1002/jobm.201800195 30198089 2-s2.0-85053311636 12 58 1100 1104 en Journal of Basic Microbiology © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. All rights reserved. |
| spellingShingle | Engineering::Bioengineering CRISPR/Cpf1 Large Fragment Deletion Li, Zhen-Hai Liu, Min Lyu, Xiao-Mei Wang, Feng-Qing Wei, Dong-Zhi CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae |
| title | CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae |
| title_full | CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae |
| title_fullStr | CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae |
| title_full_unstemmed | CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae |
| title_short | CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae |
| title_sort | crispr cpf1 facilitated large fragment deletion in saccharomyces cerevisiae |
| topic | Engineering::Bioengineering CRISPR/Cpf1 Large Fragment Deletion |
| url | https://hdl.handle.net/10356/139529 |
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