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相关概念视频

Allosteric Proteins-ATCase01:19

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Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
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CRISPR/Cas9 Genome Editing01:28

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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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Allosteric Regulation01:08

Allosteric Regulation

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Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...
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Ligand Binding and Linkage00:49

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Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked.  In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...
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Conservative Site-specific Recombination and Phase Variation02:53

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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.
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CRISPR01:59

CRISPR

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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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对Cas9特异性的差异性Allosteric调制

Yuanhao Li1, Xin Li1, Yingjie Chen1

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

Journal of chemical theory and computation
|January 9, 2026
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概括

通过改变导向RNA (gRNA) 或Cas9.9来提高CRISPR-Cas9基因编辑精度. 这些变化虽然远离活性部位,但通过固定HNH核酶域来控制Cas9,从而增强编辑特异性.

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科学领域:

  • 分子生物学分子生物学
  • 生物物理学的生物物理.
  • 基因组学就是基因组学.

背景情况:

  • 克里斯普尔-Cas9基因编辑依赖于精确的准,但偏离目标的切割仍然是一个挑战.
  • 修改导向RNA (gRNA) 和Cas9变体等策略旨在提高特异性.
  • 远程修饰如何影响Cas9活性的基础分子机制尚未完全理解.

研究的目的:

  • 阐明PAM远端变化调节CRISPR-Cas9活性和特异性的分子机制.
  • 调查gRNA截断,基不匹配和Cas9突变如何影响Cas9的结构动力学和全调节.
  • 建立一个动态框架,将远程结构干扰与Cas9活动控制联系起来.

主要方法:

  • 使用了近毫秒的全原子分子动力学模拟.
  • 模拟分析了各种PAM远程扰动对Cas9的影响.
  • 研究了Cas9的形态动态和性调节.

主要成果:

  • 所有测试的扰动,包括gRNA截断和基不匹配,通过改变HNH动态来阻碍过渡到催化活性状态.
  • 对于不同的干扰,确定了不同的全路径,包括REC3重定位和L2链接器参与.
  • 发现进化的突变重塑了全球运动模式,限制了HNH的灵活性.

结论:

  • 多个结构途径汇聚在HNH固定和催化抑制上,统一了RNA,DNA和蛋白质修饰的影响.
  • 这项研究提供了对Cas9忠诚度调节的机制性见解.
  • 这些发现为设计下一代高精度基因组编辑器提供了原则.