检索结果-南通市图书馆

Large chromosomal segment deletions by CRISPR/LbCpf1-mediated multiplex gene editing in soybean

Journal of Integrative Plant Biology 2021年第9期63卷 1620-1631页

作者： Kaixuan Duan Yuanyuan Cheng Jing Ji Chenchen Wang Yongshu Wei Yuanchao Wang Department of Plant Pathology Nanjing Agricultural UniversityNanjing 210095China

The creation of new soybean varieties has been limited by genomic duplication and redundancy.Efficient multiplex gene editing and large chromosomal segment deletion through clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein(Cas)systems are promising strategies for overcoming these obstacles.CRISPR/Cpf1 is a robust tool for multiplex gene editing.However,large chromosomal excision mediated by CRISPR/Cpf1 has been reported in only a few non-plant species.Here,we report on CRISPR/LbCpf1-induced large chromosomal segment deletions in soybean using multiplex gene targeting.The CRISPR/LbCpf1 system was optimized for direct repeat and guide RNA lengths in crispr RNA(crRNA)array.The editing efficiency was evaluated using LbCpf1 driven by the CaMV35S and soybean ubiquitin promoter.The optimized system exhibited editing efficiencies of up to 91.7%.Our results showed eight gene targets could be edited simultaneously in one step when a single eight-gRNA-target crRNA array was employed,with an efficiency of up to 17.1%.We successfully employed CRISPR/LbCpf1 to produce small fragments(gene clusters in soybean.Together,these data demonstrate the power of the CRISPR/LbCpf1 platform for multiplex gene editing and chromosomal segment deletion in soybean,supporting the use of this technology in both basic research and agricultural applications.

关键词： chromosomal segment deletion CRISPR/LbCpf1 multiplex gene editing soybean

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

A cluster of mutagenesis revealed an osmotic regulatory role of the OsPIP1 genes in enhancing rice salt tolerance

引用

The Crop Journal 2023年第4期11卷 1204-1217页

作者： Leyuan Tao Bing Wang Shichao Xin Wei Li Shengcai Huang Laihua Liu Jing Cui Qianru Zhang Xianguo Cheng State Key Laboratory of Efficient Utilization of Arid and Semi-arid Arable Land in Northern China Institute of Agricultural Resources and Regional PlanningChinese Academy of Agricultural SciencesBeijing 100081China Department of Plant Nutrition College of Resources and Environmental SciencesChina Agricultural UniversityBeijing 100193China Key Laboratory of Rubber Biology and genetic Resources of the Ministry of Agriculture Rubber Research InstituteChinese Academy of Tropical Agricultural SciencesHaikou 571011HainanChina International Joint Research Center of Technology Innovation and Achievement Transformation on Palatability of Rice Tianjin Agricultural UniversityTianjin 300384China

Aquaporins play important regulatory roles in improving plant abiotic stress tolerance.To better understand whether the Os PIP1 genes collectively dominate the osmotic regulation in rice under salt stress,a cluster editing of the Os PIP1;1,Os PIP1;2 and Os PIP1;3 genes in rice was performed by CRISPR/Cas9 system.Sequencing showed that two mutants with Cas9-free,line 14 and line 18 were successfully edited.Briefly,line 14 deleted a single C base in both the Os PIP1;1 and Os PIP1;3 genes,and inserted a single T base in the Os PIP1;2 gene,respectively.While line 18 demonstrated an insertion of a single A base in the Os PIP1;1gene and a single T base in both the Os PIP1;2 and Os PIP1;3 genes,respectively.multiplex editing of the Os PIP1 genes significantly inhibited photosynthetic rate and accumulation of compatible metabolites,but increased MDA contents and osmotic potentials in the mutants,thus delaying rice growth under salt stress.Functional loss of the Os PIP1 genes obviously suppressed the expressions of the Os PIP1,Os SOS1,Os CIPK24 and Os CBL4 genes,and increased the influxes of Na+and effluxes of K^(+)/H^(+)in the roots,thus accumulating more Na+in rice mutants under salt stress.This study suggests that the Os PIP1 genes are essential modulators collectively contributing to the enhancement of rice salt stress tolerance,and multiplex editing of the Os PIP1 genes provides insight into the osmotic regulation of the PIP genes.

关键词： Aquaporin multiplex gene editing CRISPR/Cas9 OsPIP1 genes Rice(Oryza sativa L.) Salt tolerance

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Highly efficient multiplex base editing:One-shot deactivation of eight genes in Shewanella oneidensis MR-1

引用

Synthetic and Systems Biotechnology 2023年第1期8卷 1-10页

作者： Yaru Chen Meijie Cheng Yan Li Lin Wang Lixia Fang Yingxiu Cao Hao Song Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering(Ministry of Education) School of Chemical Engineering and TechnologyTianjin UniversityTianjin300072China Key Laboratory of Systems Bioengineering(Ministry of Education) Tianjin UniversityTianjin300072China

Obtaining electroactive microbes capable of efficient extracellular electron transfer is a large undertaking for the scalability of bio-electrochemical systems.Inevitably,researchers need to pursue the co-modification of multiple genes rather than expecting that modification of a single gene would make a significant contribution to improving extracellular electron transfer rates.Base editing has enabled highly-efficient gene deactivation in model electroactive microbe Shewanella oneidensis MR-1.Since multiplexed application of base editing is still limited by its low throughput procedure,we thus here develop a rapid and efficient multiplex base editing system in S.oneidensis.Four approaches to express multiple gRNAs were assessed firstly,and transcription of each gRNA cassette into a monocistronic unit was validated as a more favorable option than transcription of multiple gRNAs into a polycistronic cluster.Then,a smart scheme was designed to deliver one-pot assembly of multiple gRNAs.3,5,and 8 genes were deactivated using this system with editing efficiency of 83.3%,100%and 12.5%,respectively.To offer some nonrepetitive components as alternatives genetic parts of sgRNA cassette,different promoters,handles,and terminators were screened.This multiplex base editing tool was finally adopted to simultaneously deactivate eight genes that were identified as significantly downregulated targets in transcriptome analysis of riboflavin-overproducing strain and control strain.The maximum power density of the multiplex engineered strain HRF(8BE)in microbial fuel cells was 1108.1 mW/m2,which was 21.67 times higher than that of the wild-type strain.This highly efficient multiplexed base editing tool elevates our ability of genome manipulation and combinatorial engineering in Shewanella,and may provide valuable insights in fundamental and applied research of extracellular electron transfer.

关键词： multiplex gene editing Electroactive microorganisms Base editing multiplexed engineering CRISPR Extracellular electron transfer

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：