相关实验视频

Updated: Jun 14, 2025

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

一个改进的CRISPR和CRISPR干扰 (CRISPRi) 工具包,用于设计模型甲原体archaeon Methanococcus maripaludis

Qing Du^1,2, Yufei Wei^1,3, Liuyang Zhang¹

¹State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No.1 Beichen West Road, Beijing, 100101, China.

Microbial cell factories

|September 3, 2024

概括

研究人员对Methanococcus maripaludis改进了CRISPR-Cas9工具,从而实现了高效的基因组编辑和基因调节. 这项工作增强了它作为生物技术和基础研究的考古模型的使用.

科学领域:

微生物学微生物学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

在古物中,CRISPR-Cas系统未得到充分利用.
甲球菌 (Methanococcus maripaludis) 是考古生物学和基于二氧化碳的生物技术中的一个关键模型生物.
有限的遗传工具阻碍了M. maripaludis的应用.

研究的目的:

加强CRISPR-Cas9工具用于M. maripaludis的基因组编辑.
开发一种用于控制基因调节的CRISPR干扰 (CRISPRi) 系统.
为了扩大M. maripaludis作为一个模型的考古细胞工厂.

主要方法:

开发了多个单导向RNAs (sgRNAs) 的平衡表达策略.
设计了一种表达Cas9的菌株,以简化等离子体结构和高效的基因敲进.
建立了使用催化无活性dCas9进行基因抑制的CRISPR干扰 (CRISPRi) 系统.
使用TetR/tetO平台创建了一个可诱导的CRISPRi-dCas9系统.

主要成果:

实现了高效的多重基因组编辑和CRISPR等离子体构造.
启用了无标记物和无痕基因敲进.
通过使用CRISPRi.证明了高达100倍的基因抑制.
成功实施诱导性基因抑制,特别是对于必不可少的基因.

结论:

扩展了M. maripaludis. 的遗传操纵工具包.
弥合了受控基因调节的方法差距,特别是对于必需的基因.
为M. maripaludis铺平了道路,成为研究和生物技术的重要考古细胞工厂.

关键词:

甲球菌 (Methanococcus maripaludis) 是一种可怕的菌株.考古物 (Archaea) 是一种古老的物种.这就是CRISPR干扰的原因.这是一个CRISPR工具箱.基因表达基因表达基因表达甲基因的产生.多重基因组编辑多重基因组编辑

更多相关视频

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

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

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...

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...