Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicum
Abstract Corynebacterium glutamicum is widely used as microbial cell factory for various bioproducts, but its genomic editing efficiency needs to be improved. In this study, a highly efficient CRISPR/Cas9-assisted genomic editing system for C. glutamicum was constructed. This system mainly involves...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
SpringerOpen
2021-05-01
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| Series: | AMB Express |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13568-021-01231-7 |
| _version_ | 1819101194326376448 |
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| author | Chengzhen Yao Xiaoqing Hu Xiaoyuan Wang |
| author_facet | Chengzhen Yao Xiaoqing Hu Xiaoyuan Wang |
| author_sort | Chengzhen Yao |
| collection | DOAJ |
| description | Abstract Corynebacterium glutamicum is widely used as microbial cell factory for various bioproducts, but its genomic editing efficiency needs to be improved. In this study, a highly efficient CRISPR/Cas9-assisted genomic editing system for C. glutamicum was constructed. This system mainly involves a plasmid and can be used for both gene insertion and deletion in the chromosome of C. glutamicum. The recombinant plasmid for the target gene containing all the editing elements, and first constructed it in E. coli, then purified and transformed it into C. glutamicum. This temperature-sensitive plasmid was cured at high temperature after the genomic editing was completed in C. glutamicum. Using this genetic editing system, the genetic editing efficiency in C. glutamicum ATCC 13032 could reach 95%. The whole work of editing could be done in 8–9 days and showed most time-saving compared to the reported. Using this system, the native promoter of gdhA1 in ATCC 13032 has been replaced with the strong promoter PtacM, and more than 10 genes in ATCC 13032 have been deleted. The results demonstrate that this CRISPR/Cas9-assisted system is highly efficient and very suitable for genome editing in C. glutamicum. |
| first_indexed | 2024-12-22T01:14:47Z |
| format | Article |
| id | doaj.art-a35ada04e90c4a5d8e88e636c9dda7bc |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-12-22T01:14:47Z |
| publishDate | 2021-05-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-a35ada04e90c4a5d8e88e636c9dda7bc2022-12-21T18:43:53ZengSpringerOpenAMB Express2191-08552021-05-0111111510.1186/s13568-021-01231-7Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicumChengzhen Yao0Xiaoqing Hu1Xiaoyuan Wang2State Key Laboratory of Food Science and Technology, Jiangnan UniversityState Key Laboratory of Food Science and Technology, Jiangnan UniversityState Key Laboratory of Food Science and Technology, Jiangnan UniversityAbstract Corynebacterium glutamicum is widely used as microbial cell factory for various bioproducts, but its genomic editing efficiency needs to be improved. In this study, a highly efficient CRISPR/Cas9-assisted genomic editing system for C. glutamicum was constructed. This system mainly involves a plasmid and can be used for both gene insertion and deletion in the chromosome of C. glutamicum. The recombinant plasmid for the target gene containing all the editing elements, and first constructed it in E. coli, then purified and transformed it into C. glutamicum. This temperature-sensitive plasmid was cured at high temperature after the genomic editing was completed in C. glutamicum. Using this genetic editing system, the genetic editing efficiency in C. glutamicum ATCC 13032 could reach 95%. The whole work of editing could be done in 8–9 days and showed most time-saving compared to the reported. Using this system, the native promoter of gdhA1 in ATCC 13032 has been replaced with the strong promoter PtacM, and more than 10 genes in ATCC 13032 have been deleted. The results demonstrate that this CRISPR/Cas9-assisted system is highly efficient and very suitable for genome editing in C. glutamicum.https://doi.org/10.1186/s13568-021-01231-7Corynebacterium glutamicumCRISPR/Cas9Metabolic engineeringGenomic editingl-Glutamic acid fermentation |
| spellingShingle | Chengzhen Yao Xiaoqing Hu Xiaoyuan Wang Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicum AMB Express Corynebacterium glutamicum CRISPR/Cas9 Metabolic engineering Genomic editing l-Glutamic acid fermentation |
| title | Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicum |
| title_full | Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicum |
| title_fullStr | Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicum |
| title_full_unstemmed | Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicum |
| title_short | Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicum |
| title_sort | construction and application of a crispr cas9 assisted genomic editing system for corynebacterium glutamicum |
| topic | Corynebacterium glutamicum CRISPR/Cas9 Metabolic engineering Genomic editing l-Glutamic acid fermentation |
| url | https://doi.org/10.1186/s13568-021-01231-7 |
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