A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy
文献类型：期刊论文
| 作者 | Wang, Jichao1; Sui, Xinyue1,3; Ding, Yamei4; Fu, Yingxin1; Feng, Xinjun1; Liu, Min1,2; Zhang, Youming2; Xian, Mo1; Zhao, Guang1,2 |
| 刊名 | NUCLEIC ACIDS RESEARCH
 |
| 出版日期 | 2021-01-25 |
| 卷号 | 49期号:2页码:15 |
| ISSN号 | 0305-1048 |
| DOI | 10.1093/nar/gkaa1141 |
| 通讯作者 | Xian, Mo(xianmo@qibebt.ac.cn) ; Zhao, Guang(zhaoguang@sdu.edu.cn) |
| 英文摘要 | The production of optimized strains of a specific phenotype requires the construction and testing of a large number of genome modifications and combinations thereof. Most bacterial iterative genome-editing methods include essential steps to eliminate selection markers, or to cure plasmids. Additionally, the presence of escapers leads to time-consuming separate single clone picking and subsequent cultivation steps. Herein, we report a genome-editing method based on a Rock-Paper-Scissors (RPS) strategy. Each of three constructed sgRNA plasmids can cure, or be cured by, the other two plasmids in the system; plasmids from a previous round of editing can be cured while the current round of editing takes place. Due to the enhanced curing efficiency and embedded double check mechanism, separate steps for plasmid curing or confirmation are not necessary, and only two times of cultivation are needed per genome-editing round. This method was successfully demonstrated in Escherichia toll and Klebsiella pneumoniae with both gene deletions and replacements. To the best of our knowledge, this is the fastest and most robust iterative genome-editing method, with the least times of cultivation decreasing the possibilities of spontaneous genome mutations. |
| 资助项目 | National Natural Science Foundation of China[31961133014] ; National Natural Science Foundation of China[31722001] ; National Natural Science Foundation of China[21807101] ; Defence Industrial Technology Development Program[JCKY2018130B005] ; Shandong University |
| WOS研究方向 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| 出版者 | OXFORD UNIV PRESS |
| WOS记录号 | WOS:000626723800006 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/170287]  |
| 专题 | 中国科学院海洋研究所 |
| 通讯作者 | Xian, Mo; Zhao, Guang |
| 作者单位 | 1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biobased Mat, Qingdao 266101, Peoples R China 2.Shandong Univ, State Key Lab Microbial Technol, Qingdao 266237, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Jichao,Sui, Xinyue,Ding, Yamei,et al. A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy[J]. NUCLEIC ACIDS RESEARCH,2021,49(2):15. |
| APA | Wang, Jichao.,Sui, Xinyue.,Ding, Yamei.,Fu, Yingxin.,Feng, Xinjun.,...&Zhao, Guang.(2021).A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy.NUCLEIC ACIDS RESEARCH,49(2),15. |
| MLA | Wang, Jichao,et al."A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy".NUCLEIC ACIDS RESEARCH 49.2(2021):15. |
入库方式： OAI收割
来源：海洋研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。