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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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CRISPR and crRNAs02:53

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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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Homologous Recombination02:31

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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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相关实验视频

Updated: Jul 24, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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一个工程化超紧的CRISPR-Cas12f系统,增强了基因编辑活动.

Tong Wu1,2, Chang Liu1, Siyuan Zou1,2

  • 1Department of Chemistry, The University of Chicago, Chicago, IL, USA.

Nature chemical biology
|July 3, 2023
PubMed
概括

研究人员设计了一个更小,更强大的CRISPR-Cas酶,enAsCas12f,用于精确的基因编辑. 这种增强的系统为遗传疾病治疗提供了改进的DNA裂变活性和最小的非目标效应.

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科学领域:

  • 分子生物学分子生物学
  • 生物技术是生物技术.
  • 遗传学 是一个遗传学.

背景情况:

  • 克里斯普尔-卡斯系统是基因编辑的强大工具.
  • 紧的CRISPR-Cas系统对于治疗应用是可取的.
  • 现有的系统通常在编辑活动和大小方面存在局限性.

研究的目的:

  • 为了设计一种更强大,更紧的CRISPR-Cas酶.
  • 评估工程系统的基因编辑效率和特异性.
  • 阐明酶活动的结构基础.

主要方法:

  • 蛋白质工程的AsCas12f,以创建一个enAsCas12f.
  • 在体外DNA裂变测试.
  • 在人类细胞中进行基因编辑实验.
  • 电子显微镜 (cryo-EM) 结构分析.
  • 单导向RNA (sgRNA) 的工程.

主要成果:

  • 工程设计的enAsCas12f比野生类型的AsCas12f.高出11.3倍的功效.
  • enAsCas12f在人体细胞中具有广泛的功能,可实现高达69.8%的插入和删除.
  • 观察到高目标活动与最小的目标外编辑.
  • 低温EM结构显示了二分化介导的基质识别.
  • 改造的sgRNA-v2较短,具有可比活动.

结论:

  • 设计的超紧的AsCas12f系统使得基因编辑功能强大且可靠.
  • enAsCas12f为遗传疾病治疗提供了一个有前途的工具.
  • 结构洞察力指导进一步优化CRISPR-Cas系统.