An improved plant CRISPR-Cas9 system for generating Cas9-free multiplex mutants
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas9 mediated gene editing systems have been used in a variety of organisms, including plants. This system can efficiently edit target genes to generate mutants, especially multiplex mutants, which are invaluable genetic materials fo...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2021-01-01
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| Series: | Biotechnology & Biotechnological Equipment |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/13102818.2021.1986132 |
| _version_ | 1798014643422101504 |
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| author | Jun-Li Wang Luo-Yu Liang Nan Lang Yu-Yang Li Wang Guo Jian Cui Ya-Jie Zou Hai-Qin Liu Qiong-Hui Fei Xiao-Feng Li Guang-Qin Guo Lei Wu |
| author_facet | Jun-Li Wang Luo-Yu Liang Nan Lang Yu-Yang Li Wang Guo Jian Cui Ya-Jie Zou Hai-Qin Liu Qiong-Hui Fei Xiao-Feng Li Guang-Qin Guo Lei Wu |
| author_sort | Jun-Li Wang |
| collection | DOAJ |
| description | CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas9 mediated gene editing systems have been used in a variety of organisms, including plants. This system can efficiently edit target genes to generate mutants, especially multiplex mutants, which are invaluable genetic materials for gene functional studies. Although the use of traditional hybridization technology to construct multiplex mutants has been widely used in plant research, the complex screening and identification, long construction cycle and other problems have always troubled researchers. Therefore, in the current plant science research, the use of CRISPR-Cas9 mediated multiplex genome editing to construct multiplex mutants has become a mainstream technology. Here, we designed an improved plant CRISPR-Cas9 mediated multiplex genome vector system. The In-fusion cloning technology was selected to conveniently assemble multiple tandem sgRNA (Single-guide RNA) expression frames. In this system, Cas9-free plants can be easily selected by examining mCherry fluorescence in T2 transgenic plants. By using this system, Cas9-free homozygous triple mutant was obtained, demonstrating the availability of our system. |
| first_indexed | 2024-04-11T15:21:44Z |
| format | Article |
| id | doaj.art-b8645be17bf64b438d356e6f6dd9cfd5 |
| institution | Directory Open Access Journal |
| issn | 1310-2818 1314-3530 |
| language | English |
| last_indexed | 2024-04-11T15:21:44Z |
| publishDate | 2021-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Biotechnology & Biotechnological Equipment |
| spelling | doaj.art-b8645be17bf64b438d356e6f6dd9cfd52022-12-22T04:16:21ZengTaylor & Francis GroupBiotechnology & Biotechnological Equipment1310-28181314-35302021-01-013511850185710.1080/13102818.2021.19861321986132An improved plant CRISPR-Cas9 system for generating Cas9-free multiplex mutantsJun-Li Wang0Luo-Yu Liang1Nan Lang2Yu-Yang Li3Wang Guo4Jian Cui5Ya-Jie Zou6Hai-Qin Liu7Qiong-Hui Fei8Xiao-Feng Li9Guang-Qin Guo10Lei Wu11Department of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityDepartment of MOE Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou UniversityCRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas9 mediated gene editing systems have been used in a variety of organisms, including plants. This system can efficiently edit target genes to generate mutants, especially multiplex mutants, which are invaluable genetic materials for gene functional studies. Although the use of traditional hybridization technology to construct multiplex mutants has been widely used in plant research, the complex screening and identification, long construction cycle and other problems have always troubled researchers. Therefore, in the current plant science research, the use of CRISPR-Cas9 mediated multiplex genome editing to construct multiplex mutants has become a mainstream technology. Here, we designed an improved plant CRISPR-Cas9 mediated multiplex genome vector system. The In-fusion cloning technology was selected to conveniently assemble multiple tandem sgRNA (Single-guide RNA) expression frames. In this system, Cas9-free plants can be easily selected by examining mCherry fluorescence in T2 transgenic plants. By using this system, Cas9-free homozygous triple mutant was obtained, demonstrating the availability of our system.http://dx.doi.org/10.1080/13102818.2021.1986132crispr-cas9multiplex genome editingin-fusion cloning technology cas9-free |
| spellingShingle | Jun-Li Wang Luo-Yu Liang Nan Lang Yu-Yang Li Wang Guo Jian Cui Ya-Jie Zou Hai-Qin Liu Qiong-Hui Fei Xiao-Feng Li Guang-Qin Guo Lei Wu An improved plant CRISPR-Cas9 system for generating Cas9-free multiplex mutants Biotechnology & Biotechnological Equipment crispr-cas9 multiplex genome editing in-fusion cloning technology cas9-free |
| title | An improved plant CRISPR-Cas9 system for generating Cas9-free multiplex mutants |
| title_full | An improved plant CRISPR-Cas9 system for generating Cas9-free multiplex mutants |
| title_fullStr | An improved plant CRISPR-Cas9 system for generating Cas9-free multiplex mutants |
| title_full_unstemmed | An improved plant CRISPR-Cas9 system for generating Cas9-free multiplex mutants |
| title_short | An improved plant CRISPR-Cas9 system for generating Cas9-free multiplex mutants |
| title_sort | improved plant crispr cas9 system for generating cas9 free multiplex mutants |
| topic | crispr-cas9 multiplex genome editing in-fusion cloning technology cas9-free |
| url | http://dx.doi.org/10.1080/13102818.2021.1986132 |
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