相关实验视频
Updated: Jan 25, 2026

08:20
Genome Engineering of Primary Human B Cells Using CRISPR/Cas9
Published on: November 3, 2020
6.9K
克里斯普尔-卡斯9跨裂变受到侧面R循环,延长的间隔器和非活跃的HNH域的阻碍
Roser Montagud-Martínez1, Raúl Ruiz1, Sara Baldanta1
1Institute for Integrative Systems Biology (I2SysBio), CSIC - University of Valencia, Paterna, Spain.
Nature communications
|January 23, 2026
概括
CRISPR Cas9酶可以切割单链DNA. 它的RNA导向DNA处理受R环,导向RNA长度和特定的Cas9域的影响,允许其功能进行微调.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 基因编辑技术的技术
背景情况:
- 克里斯普尔-卡斯9系统是基因组编辑的强大工具.
- 了解Cas9活动的精确机制对于其有效应用至关重要.
- Cas9表现出超出其预期的DNA准功能的附带活动.
研究的目的:
- 阐明控制Cas9.9通过RNA引导单链DNA处理的关键因素.
- 为了研究R环结构对Cas9的跨裂变活性的影响.
- 确定引导RNA间隔长度和Cas9域在调节Cas9功能的作用.
主要方法:
- 生物化学测试以评估Cas9在单链DNA上的活性.
- 通过RNA引导的DNA裂变实验,使用不同的R循环配置.
- 位点定向的突变发生,以探测Cas9域 (RuvC和HNH) 的功能.
- 分析结构模型以推断机械细节.
主要成果:
- 在指导RNA的5'侧的非侧面R环促进Cas9跨裂变活性,特别是在短双链DNA点上.
- 导向RNA间隔器延长超过20个基显著抑制了这种附带裂解活动.
- 虽然RuvC域对于跨裂变至关重要,但具有催化活性的HNH域提高了该过程的效率.
- 结构建模为这些观测提供了初步的机制性见解.
结论:
- Cas9的附带单链DNA处理是由目标DNA和导向RNA的特定结构特征调节的.
- Cas9的RuvC和HNH域在调解和增强跨裂变活动方面发挥着不同的作用.
- 这些发现为微调Cas9功能提供了基础,通过精确控制其分子上下文和域活性来微调Cas9功能.
相关概念视频
CRISPR and crRNAs
18.8K
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...
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...
18.8K
CRISPR
57.6K
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...
57.6K
CRISPR/Cas9 Genome Editing
1.8K
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...
1.8K
Transcription Elongation Factors
13.5K
Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA...
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA...
13.5K
Transcription Elongation Factors
4.6K
4.6K
Conservation of Protein Domains Over Different Proteins
14.1K
Protein domains are small structurally independent units that are part of a single amino acid chain. Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
14.1K

