相关实验视频
Updated: Feb 19, 2026

07:56
Genome Editing in Mammalian Cell Lines using CRISPR-Cas
Published on: April 11, 2019
23.3K
在CRISPR-Cas适应过程中对Cas9介导的预测器选择的结构性见解
Ugne Gaizauskaite1, Giedre Tamulaitiene1, Arunas Silanskas1
1Institute of Biotechnology, Life Sciences Center, Vilnius University, Saulėtekio av. 7, 10257 Vilnius, Lithuania.
Molecular cell
|February 17, 2026
概括
克里斯普尔-卡斯适应涉及卡斯9蛋白在选择免疫的外来DNA. 结构研究揭示了Cas9,Cas1-Cas2和Csn2如何形成一个超级复合体,以调解Streptococcus thermophilus中的预空间选择.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 结构生物学 结构生物学
背景情况:
- 克里斯普尔-卡斯系统在原核生物中提供适应性免疫力.
- 间隔器获取,这是一个关键的适应步骤,涉及将外来DNA集成到CRISPR阵列中.
- Cas1-Cas2整合酶和辅助蛋白质调解了间隔器的获取,在II-A型系统中发现了Cas9的参与.
研究的目的:
- 阐明在Streptococcus thermophilus II-A型CRISPR-Cas系统中Cas9介导的预位子选择的结构机制.
- 描述预测器选择超级复杂的组成和构造.
主要方法:
- 使用冷电子显微镜 (cryo-EM) 来确定高分辨率结构.
- 结构分析的重点是Streptococcus thermophilus II-A类型的超级复合物.
主要成果:
- 在DNA扫描中,超级复杂的结构和两个明显的原始空间体相邻动机 (PAM) 结合状态得到了解决.
- 这些发现为Cas9在适应过程中如何指导特定DNA序列的选择提供了原子层面的见解.
- 这项研究强调了Cas9-Cas1-Cas2-Csn2超复合体的形成.
结论:
- Cas9通过与整合酶和辅助蛋白形成超级复合体,在预位元选择中发挥着至关重要的作用.
- 结构数据揭示了CRISPR-Cas适应过程中Cas9介导的DNA标选择的机制.
- 这项工作证明了Cas9的可塑性,并扩大了对其生物功能的理解,超越了基因编辑.
相关概念视频
CRISPR/Cas9 Genome Editing
2.0K
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...
2.0K
CRISPR
58.1K
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...
58.1K
CRISPR and crRNAs
19.3K
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...
19.3K
The Antiviral System of Bacteria and Archaea: CRISPR
767
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...
767
Homologous Recombination
63.9K
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...
63.9K
Conservative Site-specific Recombination and Phase Variation
6.9K
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...
6.9K

