相关实验视频
Updated: Mar 31, 2026

11:53
Substrate Generation for Endonucleases of CRISPR/Cas Systems
Published on: September 8, 2012
28.2K
在CRISPR-Cas系统中采用PAM依赖间隔器的结构和机制基础
Jiuyu Wang1, Jiazhi Li2, Hongtu Zhao2
1Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; Beijing Key Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
Cell
|October 20, 2015
概括
细菌使用CRISPR-Cas系统来记住病毒DNA. 这项研究揭示了Cas1-Cas2复合体的结构,展示了它们如何选择并将病毒DNA片段 (间隔器) 插入CRISPR阵列以进行免疫记忆.
科学领域:
- 微生物学
- 分子生物学
- 结构生物学
背景情况:
- 细菌通过CRISPR-Cas系统对病毒具有适应性免疫力.
- CRISPR 阵列将来自入侵的菌体和等离子体的 DNA 序列存储为间隔器.
- 获得隔离器是建立适应性免疫力的关键步骤.
研究的目的:
- 阐明Cas1-Cas2复合体对原空间细胞选择和DNA处理的结构机制.
- 了解Protospacer相邻基因 (PAM) 在CRISPR-Cas适应性免疫中的作用.
主要方法:
- 使用X射线结晶学来确定Cas1-Cas2-DNA复合物的结构.
- 进行生物化学测试以分析DNA结合和分裂活动.
主要成果:
- 这项研究揭示了Cas1-Cas2复合物的结构与侧面3'悬架的分叉原体空间DNA结合在一起.
- Cas1a子单元以特定的基础方式识别3'悬浮中的PAM序列.
- 从双重边界发生5nt的裂变,产生33nt的插入中间体.
- 细胞间隔剂结合会诱导Cas1-Cas2的形状变化,从而促进识别.
结论:
- 这些发现提供了基于结构的机械洞察力,以了解PAM依赖的原始空间器的获取过程.
- 这项工作阐明了CRISPR-Cas系统如何采样和处理外来DNA以获得适应性免疫力.
相关概念视频
The Antiviral System of Bacteria and Archaea: CRISPR
990
CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
990
CRISPR and crRNAs
19.5K
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...
19.5K
CRISPR
59.2K
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...
59.2K
CRISPR
18.9K
18.9K
CRISPR/Cas9 Genome Editing
2.7K
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...
2.7K
Conservative Site-specific Recombination and Phase Variation
7.3K
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...
The recognition sites for Cre recombinase called LoxP...
7.3K

