相关实验视频

Updated: Jul 16, 2025

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

Published on: December 11, 2020

通过条件稳定CRISPR-Cas13b编辑器进行删除

Ying Xu¹, Yufan Wang¹, Fu-Sen Liang¹

¹Department of Chemistry, Case Western Reserve University, 2080 Adelbert Rd, Cleveland, OH 44106, USA.

Angewandte Chemie (International ed. in English)

|September 15, 2023

概括

研究人员开发了一种条件RNA编辑平台,使用联体稳定dCas13编辑器进行精确的N6-甲基氨酸 (m⁶ A) 修改. 这种系统允许在m⁶ A上进行可诱导控制,从而最大限度地减少功能研究的固态干扰.

科学领域:

分子生物学分子生物学
RNA表观遗传学在RNA表观遗传学.
基因编辑技术的技术

背景情况:

N6-甲基氨酸 (m⁶ A) 是一种关键的RNA修饰,可以调节各种生物过程.
克里斯普尔技术能够进行可编程的编辑,但蛋白质的大小和构成性表达带来了挑战.
现有的CRISPR/RNA编辑酶可以干扰原生RNA和细胞功能.

研究的目的:

开发一个有条件和带诱导的平台,用于可编程的m⁶ A编辑.
为了克服大型CRISPR蛋白质和RNA编辑中的构成性酶表达的局限性.
为了使特定站点的m⁶ 能够在功能研究中以最小的硬体干扰进行擦除.

主要方法:

开发一个有条件的m6编辑平台 (FKBP*-dCas13b-ALK) 使用一个联体稳定dCas13编辑器.
m6的诱导表达通过添加或删除Shield-1分子控制的编辑系统.
针对dCas13b-m⁶的有针对性的招募的演示,一个擦拭器融合蛋白和特定地点的m⁶在Shield-1控制下的擦除.

主要成果:

一个有条件的,联体诱导的编辑系统的成功实施.
盾-1分子有效地控制了dCas13b融合蛋白的表达和招募,以向目标m6A删除.
在Shield-1被移除后,dCas13b融合蛋白的快速释放和降解,在恢复之前.

结论:

FKBP*-dCas13b-ALK平台提供了一种新的方法,用于条件和可诱导的编辑.
这个系统允许精确控制m⁶A的删除,促进研究m⁶A的功能.
融合蛋白的快速清除将固体障碍降到最低,为研究RNA修饰作用提供了改进的方法.

关键词:

在Cas13b中使用.蛋白质降解蛋白质降解这是一个RNARNARNARNARNA.盾牌-1 盾牌-1 是一个 m6A 一个很好的.

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

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

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...

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...