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CRISPR01:59

CRISPR

50.8K
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...
50.8K
CRISPR and crRNAs02:53

CRISPR and crRNAs

17.0K
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...
17.0K
Homologous Recombination02:31

Homologous Recombination

50.5K
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...
50.5K
The Spindle Assembly Checkpoint02:19

The Spindle Assembly Checkpoint

3.2K
The spindle assembly checkpoint is a molecular surveillance mechanism ensuring the fidelity of chromosome segregation during anaphase. The checkpoint monitors the completion of all the prerequisite steps before chromosome segregation to determine whether the segregation process should proceed or be delayed.
Many proteins function together to control the spindle assembly checkpoint. Mutations affecting these proteins may allow cells to proceed into anaphase prematurely, resulting in the...
3.2K

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相关实验视频

Updated: Jun 28, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

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在CRISPR-Cas9中使用二次合规检查点.

Shuxin Zhao1,2, Jin Liu3, Zhicheng Zuo1,2

  • 1College of Chemistry and Chemical Engineering, Shanghai University of Engineering Science, Shanghai 201620, China.

Journal of chemical theory and computation
|April 16, 2024
PubMed
概括

研究人员在Cas9基因编辑中发现了一种二次检查点,该检查点可以提高精度. 进化的Cas9变种加强了这个检查点,通过控制DNA裂变激活来提高精度.

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相关实验视频

Last Updated: Jun 28, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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科学领域:

  • 分子生物学分子生物学
  • 生物化学 生物化学
  • 遗传学 遗传学 是一个

背景情况:

  • CRISPR-Cas9基因编辑精度至关重要,但在分子层面上还没有完全理解.
  • 现有的CRISPR-Cas9变体旨在提高DNA裂变的准确性,但它们的机制需要更深入的研究.

研究的目的:

  • 调查Cas9的后期结构动力学和在分离前状态中的一个进化变体 (evoCas9).
  • 阐明 evoCas9.9 的增强精度背后的分子机制.

主要方法:

  • 用次毫秒的动态模拟来观察形状转变.
  • 分析的重点是HNH核酶域的运动和全性通信网络.

主要成果:

  • 基因不匹配破坏了Cas9的HNH核酶域运动,损害了 conformational 激活,并暗示了一个次要检查点.
  • evoCas9表现出改变的激活路径与不匹配,包括HNH域重新定位和扰乱的全网络.
  • 在evoCas9中发生的突变加强了初级和二级检查点,确保了高基因编辑精度.

结论:

  • 一个二次构造检查点微调Cas9DNA裂变激活,特别是在存在基因不匹配的情况下.
  • 进化的Cas9变种 (evoCas9) 使用增强的检查点机制,以获得更高的基因编辑效率.
  • 鉴定的机制可能适用于其他高保真 Cas9 导向RNA 变体.