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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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...
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CRISPR01:59

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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...
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Homologous Recombination02:31

Homologous Recombination

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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...
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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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.
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Caspases01:24

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Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
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相关实验视频

Updated: Aug 23, 2025

CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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CIRCLE-Seq for Interrogation of Off-Target Gene Editing

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针对Cas9非目标活动的结构基础

Martin Pacesa1, Chun-Han Lin2, Antoine Cléry3

  • 1Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.

Cell
|October 28, 2022
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概括

CRISPR-Cas9基因组编辑可以针对意想不到的DNA序列,这引发了安全问题. 结构分析显示非正规的基配对和删除的容纳使得目标外结合,指导改进的导向RNA设计.

关键词:
关于CRISPR的研究其他国家射线晶体学基础配对基因组编辑导向RNA没有匹配中核酶在目标之外

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科学领域:

  • 分子生物学
  • 遗传学
  • 生物化学

背景情况:

  • 与CRISPR相关的 (Cas) 核酶Cas9是一种强大的基因组编辑工具.
  • Cas9的特异性依赖于指导RNA与向DNA的互补性.
  • 在临床应用中,Cas9的脱会带来安全风险.

研究的目的:

  • 阐明Cas9脱和裂变的结构基础.
  • 了解Cas9如何适应目标DNA中的不匹配和删除.
  • 为了合理设计更安全的基于Cas9的基因组编辑系统.

主要方法:

  • 对Cas9DNA复合物的X射线晶体学.
  • 对不同互补性的非目标基质进行分析.
  • 在目标和目标以外的绑定模式的结构比较.

主要成果:

  • Cas9通过非正规的基配对相互作用来结合非目标DNA.
  • 单核酸删除通过基跳转或多个非正规对进行.
  • PAM-远距离不匹配导致Cas9的双重脱配和形状变化.

结论:

  • 结构洞察力解释了Cas9的目标外活动.
  • 这些发现有助于改进指导RNA设计以提高特异性.
  • 这项工作有助于为CRISPR技术开发更好的目标外预测算法.