相关实验视频

Updated: Jun 25, 2025

Cell-free Protein Expression Using the Rapidly Growing Bacterium Vibrio natriegens

Cell-free Protein Expression Using the Rapidly Growing Bacterium Vibrio natriegens

Published on: March 14, 2019

移动CRISPRi作为调节Vibrio基因表达的强大工具

Logan J Geyman¹, Madeline P Tanner², Natalia Rosario-Meléndez^3,4

¹Department of Biology, Indiana University, Bloomington, Indiana, USA.

Applied and environmental microbiology

|May 22, 2024

概括

移动CRISPRi系统能够在五种Vibrio物种中进行强大的基因敲除,从而促进对基本和非基本基因的研究. 这种工具对于理解Vibrio至关重要.

科学领域:

微生物学微生物学
分子生物学分子生物学
遗传学遗传学是一种遗传学.

背景情况:

克里斯普尔干扰 (CRISPRi) 是一种强大的基因淘汰技术,利用失活的Cas9 (dCas9) 和引导RNA (sgRNA) 来阻止转录.
移动CRISPRi提供了一个模块化载体系统,通过Tn7转换将dCas9和sgRNA集成到细菌基因组中,从而实现可诱导的基因沉默.
活菌物种是重要的海洋居民,具有生态和人类健康的影响,需要有效的遗传工具来研究它们.

研究的目的:

为了证明移动CRISPRi系统在多种Vibrio物种中对基因敲除的有效性和特异性.
验证系统能够针对基本基因和非基本基因的能力,从而导致可观测的表型.
建立移动CRISPRi作为一个功能基因组学在Vibrio的多功能工具.

主要方法:

使用Tn7转换将IPTG诱导的dcas9和sgRNA基因集成到五种Vibrio物种的基因组中.
设计和利用针对特定基本基因和非基本基因的sgRNA.
通过观察由此产生的表型,如生物发光,定量感应,细胞分裂和生长停止等来评估基因淘汰效率.

主要成果:

移动CRISPRi系统在五种Vibrio物种中展示了强大的和特定的基因淘汰:V. cholerae,V. fischeri,V. vulnificus,V. parahaemolyticus和V. campbellii.
成功准参与生物发光,定量感应,细胞分裂和生长停止的基因,导致了清晰,可测量的表型.
该系统证明对基本基因和非基本基因都有效,证实了它在Vibrio属内的广泛适用性.

结论:

移动CRISPRi是一种高效和适应性强的工具,用于对各种维氏体物种进行基因操纵.
这种系统使研究人员能够系统地研究基因功能和本质性,从而推进我们对菌生物学的理解.
移动CRISPRi在Vibrio物种中的强大性能为研究它们在海洋生态系统中的作用及其对人类健康的影响开辟了新的途径.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?克里斯普里是什么?克里斯普里是什么?这就是Vibrio Vibrio.维布里奥·坎贝利 (Vibrio campbellii) 是一个霍乱病毒病毒 (Vibrio cholerae) 是一种病毒.维布里奥·菲舍里 (Vibrio fischeri) 是一种鱼类.这种病毒是Vibrio parahaemolyticus.这种病毒名为Vibrio vulnificus.基因表达的基因表达方式

更多相关视频

TransFLP — A Method to Genetically Modify Vibrio cholerae Based on Natural Transformation and FLP-recombination

TransFLP — A Method to Genetically Modify Vibrio cholerae Based on Natural Transformation and FLP-recombination

Published on: October 8, 2012

Author Spotlight: Advancing Therapeutics to Treat Vibriosis in Humans and Aquatic Organisms

Author Spotlight: Advancing Therapeutics to Treat Vibriosis in Humans and Aquatic Organisms

Published on: May 31, 2024

相关概念视频

Riboswitches

Riboswitches

Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...

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...

Bacterial Transcription

Bacterial Transcription

RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:

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...

Transcription Attenuation in Prokaryotes

Transcription Attenuation in Prokaryotes

Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure. Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...

Bacterial Signaling

Bacterial Signaling

Bacterial signaling can occur within bacteria (intracellular) or between bacteria (intercellular). At times, a group of bacteria behaves like a community. To achieve this, they engage in quorum sensing, the perception of higher cell density that causes changes in gene expression. Quorum sensing involves both extracellular and intracellular signaling. The signaling cascade starts with a molecule called an autoinducer (AI). Individual bacteria produce AIs that move out of the bacterial cell...