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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

211
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

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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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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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: Sep 10, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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为高效多重基因组编辑而设计的CRISPR-Cas12i工具

Linli Wang1,2,3, Yanlu Wang1,2,3, Jian Chen4

  • 1State Key Laboratory of Animal Biotech Breeding, China Agricultural University, Beijing 100193, China.

Nucleic acids research
|August 28, 2025
PubMed
概括

研究人员设计了CRISPR RNA (crRNA) 导向核酶Cas12i.3,以改善多重基因组编辑. 增强的优化Cas12i (EOCas12i) 系统可以同时有效编辑多个目标,为遗传研究提供了简化工具.

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科学领域:

  • 分子生物学
  • 基因组工程
  • 生物技术

背景情况:

  • 多重复合基因组编辑工具在前体CRISPRRNA (crRNA前) 处理方面面临挑战,需要额外的调控组件.
  • Cas12i.3核酶缺乏crRNA前处理能力,限制了其复合潜力.

研究的目的:

  • 设计基于Cas12i.3的高效和简单的多重基因组编辑系统.
  • 克服野生类型Cas12i.3在广泛基因组编辑应用中的局限性.

主要方法:

  • 优化CRISPRRNA (crRNA) 设计,编码子使用和外核酶融合以创建初始优化的Cas12i (IOCas12i).
  • 使用合理设计和氨基酸突变来开发增强优化的Cas12i (EOCas12i) 系统 (EOCas12i-Combo1和EOCas12i-Combo2).
  • 使用多达30个目标crRNA阵列评估编辑效率,特异性和多重复合能力.

主要成果:

  • 与野生型Cas12i.3相比,工程EOCas12i系统显示了显著提高的编辑效率 (2,5至60倍).
  • 编辑效率与已建立的系统如Streptococcus pyogenes Cas9 (SpCas9) 和Lachnospiraceae细菌 Cas12a (LbCas12a) 相似.
  • 通过使用紧的crRNA阵列,对多达30个目标进行有效的多重编辑,从而产生更长的基因淘汰效果.

结论:

  • 开发的EOCas12i-Combo1和EOCas12i-Combo2系统代表了多重基因组编辑的重大进步.
  • 这些工程核酶为各种基因组编辑应用提供了简化和高效的平台.
  • 增强的Cas12i变种克服了以前的局限性,为在基因研究和工程中更广泛的使用铺平了道路.