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相关概念视频

CRISPR01:59

CRISPR

50.6K
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...
50.6K
Homologous Recombination02:31

Homologous Recombination

50.4K
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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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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

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相关实验视频

Updated: Jun 24, 2025

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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通过减少错误折叠的pegRNA相互作用来增强CRISPR主要编辑.

Weiting Zhang1,2, Karl Petri3,4, Junyan Ma1,2,5

  • 1Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, United States.

eLife
|June 7, 2024
PubMed
概括
此摘要是机器生成的。

在主要编辑指导RNA (pegRNAs) 中的内部序列互补性可以降低CRISPR主要编辑 (PE) 的效率. 简单的重新折叠程序和有针对性的突变显著提高了斑马鱼胚胎的PE效率.

关键词:
结构 RNA 结构 RNA 结构基因编辑 基因编辑遗传学 遗传学 遗传学 是一个基因组学就是基因组学.提供蛋白质的蛋白质输送.斑马鱼是一种斑马鱼.

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相关实验视频

Last Updated: Jun 24, 2025

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

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科学领域:

  • 分子生物学分子生物学
  • 基因编辑技术的技术
  • 生物化学 生物化学

背景情况:

  • 克里斯普尔主要编辑 (PE) 使用一个Cas9尼克酶-逆转录酶融合蛋白 (PE2) 和一个主要编辑指导RNA (pegRNA).
  • RNAs被设计为引导PE以准基因组序列并编码所需的编辑.
  • RNA结构的潜在限制可以阻碍Cas9复杂化并降低PE效率.

研究的目的:

  • 调查pEGRNA内部内部序列互补性对PE效率的影响.
  • 开发和验证改进pegRNA结构和功能的方法.
  • 为了提高CRISPR主要编辑技术的效率.

主要方法:

  • 在pegRNAs中对序列互补性的分析.
  • 开发一个PegRNA重折叠程序.
  • 引入点突变来破坏内部pegRNA相互作用.
  • 在斑马鱼胚胎中使用核蛋白复合体评估PE效率.

主要成果:

  • RNAs的5'和3'区域之间的序列互补性会对Cas9复合产生负面影响.
  • 在斑马鱼胚胎中,一个简单的pegRNA重新折叠程序将PE效率提高了25倍.
  • 引入点突变来破坏内部pegRNA相互作用进一步提高了PE效率,高达六倍.

结论:

  • 内部序列互补性是限制pegRNA疗效的关键因素.
  • 佩格RNA重新折叠和向突变发生是提高CRISPR主要编辑效率的有效策略.
  • 这些发现为优化PE用于更广泛的研究和治疗应用提供了实际方法.