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相关概念视频

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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The Nucleosome Core Particle01:12

The Nucleosome Core Particle

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Nucleosomes are the DNA-histone complex, where the DNA strand is wound around the histone core. The histone core is an octamer containing two copies of H2A, H2B, H3, and H4 histone proteins.
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their primary aim is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. On the other hand, they must allow polymerase enzymes to access histone-bound DNA during...
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Nucleosome Remodeling02:54

Nucleosome Remodeling

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Nucleosomes are the basic units of chromatin compaction. Each nucleosome consists of the DNA bound tightly around a histone core, which makes the DNA inaccessible to DNA binding proteins such as DNA polymerase and RNA polymerase. Hence, the fundamental problem is to ensure access to DNA when appropriate, despite the compact and protective chromatin structure.
Nucleosome remodeling complex
Eukaryotic cells have specialized enzymes called ATP-dependent nucleosome remodeling enzymes. These enzymes...
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The Nucleosome02:33

The Nucleosome

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DNA in a human cell is almost 2m long and it is packed inside a tiny nucleus that is only a few microns in diameter. The level of compaction of DNA inside the nucleus is astonishing. It is organized into several sequentially higher levels of compaction to fit into such a tiny space. The most compact form of DNA is a chromosome that can be seen under a microscope in a dividing cell.
DNA is wound twice around a protein complex called histone core, that consist of 8 histone proteins. This complex...
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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.
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...
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相关实验视频

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对Cas9如何向核细胞的结构洞察力.

Reina Nagamura1, Tomoya Kujirai2, Junko Kato2

  • 1Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.

Nature communications
|December 31, 2024
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概括

在CRISPR-Cas9基因组编辑中,向了真核生物中的核体DNA. Cas9优先切割链接DNA,而不是紧密包裹的DNA,为改进的基于染色体的基因编辑工具提供了洞察力.

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科学领域:

  • 分子生物学分子生物学
  • 基因组学就是基因组学.
  • 结构生物学 结构生物学

背景情况:

  • 与CRISPR相关的内核酶Cas9是一个关键的基因组编辑工具.
  • 细胞DNA被包装成染色质,核细胞作为基本单元.
  • 了解Cas9与核细胞DNA的相互作用对于真核生物基因组编辑至关重要.

研究的目的:

  • 为了研究Cas9在核体内的向的结构基础.
  • 为了阐明Cas9如何与DNA相互作用,在染色质的背景下.
  • 帮助开发先进的基因组编辑技术.

主要方法:

  • 原生聚烯胺凝电泳 (PAGE) 用于DNA裂变分析.
  • 低温电子显微镜 (cryo-EM) 用于确定Cas9-sgRNA-核体复合物的结构.
  • 在体外和体内测试以评估Cas9在核细胞DNA上的活性.

主要成果:

  • Cas9的目标是核细胞的链接DNA和入出DNA区域,避免紧密包裹的DNA.
  • 化EM揭示了Cas9和核细胞之间多个相互作用点.
  • 减少Cas9-核酶体相互作用的突变增强了体外分裂,在体内抑制有限.

结论:

  • Cas9与核细胞的相互作用会影响其DNA分裂活动.
  • 对Cas9核酶结合的结构洞察对于理解其在染色质中的功能至关重要.
  • 这些发现为设计更有效的基于染色体的基因组编辑工具铺平了道路.