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相关概念视频

CRISPR01:59

CRISPR

51.0K
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...
51.0K
CRISPR and crRNAs02:53

CRISPR and crRNAs

17.0K
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...
17.0K
Homologous Recombination02:31

Homologous Recombination

50.5K
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...
50.5K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.0K
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.
The recognition sites for Cre recombinase called LoxP...
6.0K

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相关实验视频

Updated: Jul 5, 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

Published on: May 25, 2018

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通过CRISPR-Cas12m效应器的先天可编程DNA结合,可以实现高效的基编辑.

Greta Bigelyte1, Brigita Duchovska1, Rimante Zedaveinyte1

  • 1Institute of Biotechnology, Life Sciences Center, Vilnius University, Vilnius LT-10257, Lithuania.

Nucleic acids research
|January 23, 2024
PubMed
概括

新的CRISPR-Cas12m蛋白质通过DNA结合而不是裂变提供细胞原体免疫力. 这一发现使得用于基因编辑应用的紧基基编辑工具的开发成为可能.

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科学领域:

  • 分子生物学分子生物学
  • 微生物遗传学 微生物遗传学
  • 生物化学 生物化学

背景情况:

  • 2类CRISPR-Cas系统,包括Cas9和Cas12核酶,通过通过RNA引导的外来DNA分裂,赋予 prokaryotic 免疫力.
  • 在Cas12蛋白家族中的多样性是广泛的,目前正在进行研究以表征新型亚型及其功能.

研究的目的:

  • 描述一组新型的紧型CRISPR-Cas12m (亚型V-M) 效应蛋白.
  • 阐明Cas12m蛋白在赋予 prokaryotic 免疫力的作用机制.
  • 探索Cas12m效应器在开发新基因编辑技术方面的潜力.

主要方法:

  • 生物化学测试被用来研究Cas12m效应器的功能性质.
  • 使用冷电子显微镜 (Cryo-EM) 来确定DNA结合的结构机制.
  • Cas12m与腺因除氨酶 (TadA-8e) 融合,以测试其在基编辑中的有效性.

主要成果:

  • 克里斯普尔-Cas12m蛋白质通过向的DNA结合提供对细菌菌体和质粒的保护,与DNA裂变不同.
  • Cas12m效应器作为路障,抑制DNA转录和/或复制,触发干扰.
  • 克里奥-EM结构揭示了DNA结合和干扰的分子基础,这种干扰是由Cas12m介导的.
  • GoCas12m与TadA-8e的融合导致了细菌和人类细胞中高效的A-to-G基编辑.

结论:

  • 这项研究扩大了对Cas12蛋白家族的理解,突出了Cas12m效应器中的DNA结合依赖干扰机制.
  • 这些发现揭示了一种由DNA结合而不是分裂介导的新型细胞原体免疫机制.
  • 基于Cas12m的基编辑器代表了开发紧和高效的基因编辑工具的有希望的进步.