相关实验视频

Updated: Jul 4, 2025

Designing, Packaging, and Delivery of High Titer CRISPR Retro and Lentiviruses via Stereotaxic Injection

Designing, Packaging, and Delivery of High Titer CRISPR Retro and Lentiviruses via Stereotaxic Injection

Published on: May 23, 2016

在设计菌体工程中使用Cas9准和红色再组合

Shin-Yae Choi¹, Danitza Xiomara Romero-Calle¹, Han-Gyu Cho¹

¹Program of Biopharmaceutical Science and Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Sciences, CHA University, Gyeonggi, 13488, Republic of Korea.

Journal of microbiology (Seoul, Korea)

|February 1, 2024

概括

这项研究引入了一种使用合成生物学来设计Pseudomonas aeruginosa中的细菌体 (菌素) 的新方法. 这种方法有效地为研究和治疗应用创造了设计菌体.

科学领域:

微生物学微生物学
合成生物学合成生物学
遗传学遗传学是一种遗传学.

背景情况:

菌体 (菌体) 是天然的抗菌剂和生物纳米粒子.
合成生物学工具增强了菌体应用,导致具有改进特征的设计菌体.
在Pseudomonas aeruginosa中工程菌体对于开发新型抗菌战略至关重要.

研究的目的:

开发一种简单的基因组工程方法,用于在Pseudomonas aeruginosa中创建设计菌体.
为了利用Streptococcus pyogenes Cas9 (SpCas9) 进行野生类型的菌体清除和 λRed 重组系统进行重组生成.
通过创建光菌体和评估基因组工程的效率来验证该方法.

主要方法:

通过整合SpCas9和λRed基因,构建了P. aeruginosa PAO1的生产者 (PD) 细胞.
采用了针对野生类型gp35基因的单导向RNA (sgRNA) 的等离子体和sfGFP标记的编辑模板.
通过电穿孔将等离子体引入PD细胞,以产生光菌体.

主要成果:

体基因组工程效率受到sgRNA位置和数量的影响.
双重sgRNA分子提高了菌素修饰的准效率.
从工程P. aeruginosa培养物中成功地恢复了光菌素颗粒.

结论:

建立了一种简单有效的基因组工程法菌体在P. aeruginosa的方法.
该协议有助于创建设计菌体用于研究和治疗应用.
开发的方法支持基于菌体的抗微生物策略的进步.

关键词:

针对Cas9的目标定位.设计菌体的设计者菌体基因组工程是基因组工程.重组组合是一种重组.红色的重组组合是红色的

更多相关视频

A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors

A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors

Published on: October 30, 2016

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

相关概念视频

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

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...

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...

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...