相关实验视频

Updated: May 24, 2025

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Published on: February 26, 2019

具有增强特异性的SauriCas9的工程

Xiaoqi Zhang¹, Chen Tao¹, Miaomiao Li¹

¹Center for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, School of Life Sciences, Fudan University, Shanghai 200438, China.

Molecular therapy. Nucleic acids

|March 3, 2025

概括

修改后的Cas9酶SauriCas9-R253A显著提高了基因编辑的特异性. 这种改进的变种显示出对体内治疗应用的希望,在CAR-T细胞疗法开发中没有可检测的脱效应.

科学领域:

分子生物学分子生物学
基因编辑技术的技术
生物技术是生物技术.

背景情况:

SauriCas9是一种紧的Cas9核酶,有可能用于体内治疗.
非目标效应是治疗应用中Cas9特异性的一个主要问题.
提高特异性对于安全有效地使用基因编辑工具至关重要.

研究的目的:

为了提高SauriCas9核酶的特异性.
开发一种SauriCas9变种,减少目标外影响.
为了验证增强的SauriCas9变异在CAR-T细胞疗法的基因破坏中的有效性.

主要方法:

引入了对SauriCas9的突变,以消除与目标DNA的极性接触,创建了SauriCas9-R253A变体.
利用SauriCas9-R253A破坏关键基因 (B2M,TRAC,PDCD1),这对于全基性CAR-T细胞疗法至关重要.
使用敏感检测方法评估基因破坏效率和评估非目标效应.

主要成果:

与野生类型相比,SauriCas9-R253A变种表现出增强的特异性.
使用SauriCas9-R253A.有效地破坏了B2M,TRAC和PDCD1基因.
对于具有SauriCas9-R253A.A.最有效的单导向RNA,没有观察到可检测的脱效应.

结论:

SauriCas9-R253A代表了Cas9基因编辑特异性的重大进展.
索里卡斯9-R253A的增强特异性支持其在体内安全治疗应用的潜力.
这种改进的核酶是开发下一代CAR-T细胞疗法的有希望的工具.

关键词:

这就是CRISPR-Cas.MT:RNA/DNA编辑编辑萨乌里卡斯9-R253A 类动物紧的Cas9就是一个Cas9.效率效率效率效率效率效率效率基因编辑基因编辑特殊性的特异性

更多相关视频

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

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

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...