相关实验视频

Updated: Jul 11, 2025

Generation of In-Frame Gene Deletion Mutants in Pseudomonas aeruginosa and Testing for Virulence Attenuation in a Simple Mouse Model of Infection

Generation of In-Frame Gene Deletion Mutants in Pseudomonas aeruginosa and Testing for Virulence Attenuation in a Simple Mouse Model of Infection

Published on: January 8, 2020

一种基于SEVA的CRISPR-Cas3辅助基因组工程方法,用于Pseudomonas,具有高效的载体治愈

Eveline-Marie Lammens¹, Daniel Christophe Volke², Kaat Schroven¹

¹Laboratory of Gene Technology, Department of Biosystems, KU Leuven , Leuven, Belgium.

Microbiology spectrum

|November 17, 2023

概括

这项研究引入了CRISPR-Cas3系统,以便在Pseudomonas细菌中轻松编辑基因组. 这种多功能系统还可以有效地从细菌细胞中去除不需要的载体和质粒.

科学领域:

微生物学微生物学
分子生物学分子生物学
遗传学遗传学是一个

背景情况:

克里斯普尔-卡斯系统提供精确的基因组工程能力.
开发适应性工具用于像Pseudomonas这样的阴性细菌对于合成生物学和生物技术至关重要.
有效的载体去除对于简化基因操纵工作流程至关重要.

研究的目的:

介绍一种多功能的CRISPR-Cas3编辑系统,用于Pseudomonas putida和Pseudomonas aeruginosa的基因组改变.
通过针对转移的来源来证明系统对通用向量固化的实用性.
建立一个灵活且易于转移的基因组工程平台,用于多种不同的阴性宿主.

主要方法:

使用了CRISPR-Cas3系统,提供了具有抗生素标记物的金门相容载体.
采用标准欧洲矢量架构 (SEVA) 矢量集用于同质性修复模板.
设计了一个针对向量固化应用的传输起源的间隔器.

主要成果:

在使用CRISPR-Cas3系统的Pseudomonas物种中成功创建了基因组改变.
通过针对转移的来源,在几天内有效地清除了多达三种SEVA载体.
展示了该系统的灵活性和可移植性在多个Gram-阴性主机.

结论:

提出的CRISPR-Cas3系统提供了一个简单的方法,用于编辑 Pseudomonas 的基因组.
该系统作为一种通用且有效的载体治愈工具,简化了细菌基因工程.
这种方法对推进基因组工程和在细菌中管理等离子体具有广泛的影响.

关键词:

这就是CRISPR-Cas.这就是Cas3的情况.伪omonas 是一个类型的.伪omonas aeruginosa 这种类型的病毒.伪omonas putida可以使用.塞瓦 (Sevá) 是一家国际贸易公司.基因组编辑基因组编辑矢量化矢量化

更多相关视频

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira

Published on: August 14, 2021

相关概念视频

CRISPR/Cas9 Genome Editing

CRISPR/Cas9 Genome Editing

The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...

CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...

CRISPR and crRNAs

CRISPR and crRNAs

Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...