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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

79
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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Conservative Site-specific Recombination and Phase Variation02:53

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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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相关实验视频

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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改进的原始编辑允许在Physcomitrium patens中进行常规可预测的基因编辑.

Pierre-François Perroud1, Anouchka Guyon-Debast1, Josep M Casacuberta2

  • 1Université Paris-Saclay, INRAE, AgroParisTech, Institut Jean-Pierre Bourgin (IJPB), 78000 Versailles, France.

Journal of experimental botany
|May 27, 2023
PubMed
概括

主编辑,精确的基因编辑工具,已被增强,以提高模型植物Physcomitrium patens.的效率. 这种优化的方法可以提高基因研究的编辑率,而不会影响准确度.

关键词:
菲斯科米特里姆帕斯 (Physcomitrium patens) 是一个古老的生物.这就是EpegRNA.基因组编辑 基因组编辑主编辑主要编辑.伪结是一个假结.分裂主要编辑

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科学领域:

  • 分子生物学分子生物学
  • 植物科学 植物科学
  • 遗传学 遗传学 是一个

背景情况:

  • 有效和精确的基因编辑对于逆转基因学至关重要.
  • 主编辑,CRISPR/Cas9衍生品,提供精度,但需要提高效率.

研究的目的:

  • 改进主要编辑方法,以便在Physcomitrium patens.中常规使用.
  • 调查提高主要编辑效率的策略,并探索其应用.

主要方法:

  • 在Physcomitrium patens中标准化的原始质细胞转移.
  • 评估各种主要编辑指导RNA (pegRNA) 结构和主要编辑器变体.
  • 针对APT记者基因进行直接选择和Ppdek10进行间接选择.

主要成果:

  • 优化主要编辑器表达和pegRNA修改显著增加了编辑率.
  • 反转录酶模板中的同名突变提高了效率,但没有影响编辑质量.
  • 通过间接选择证明了成功的基因编辑,并确定了用于主要编辑的植物逆转录酶.
  • 展示了使用两个独立编码的的原始编辑潜力.

结论:

  • 开发的方法允许在Physcomitrium patens.中进行常规和高效的prime编辑.
  • 在主要编辑器组件和pegRNA设计的改进是改善编辑速率的关键.
  • 主编辑是一种适用于植物直接和间接基因选择策略的多功能工具.