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相关概念视频

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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Homologous Recombination02:31

Homologous Recombination

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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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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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相关实验视频

Updated: Jun 20, 2025

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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有效的基因组编辑使用化学寡核酸导向编辑.

Long T Nguyen1,2,3, Noah R Rakestraw4, Brianna L M Pizzano5

  • 1Department of Molecular Biology, Princeton University, Princeton, NJ, USA.

bioRxiv : the preprint server for biology
|July 19, 2024
PubMed
概括
此摘要是机器生成的。

新的仿真寡核酸定向编辑 (CODE) 系统提供精确的搜索和替换基因组编辑. 这些新型编辑器在人类细胞中实现了高效的基因修改,最小的意外编辑,扩大了基因组编辑能力.

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相关实验视频

Last Updated: Jun 20, 2025

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

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科学领域:

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

背景情况:

  • 主编辑代表了精确基因组编辑的重大进步.
  • 现有的基于Cas9的方法有局限性,新的方法旨在克服这些局限性.

研究的目的:

  • 开发新的基因组编辑系统,用于精确的搜索和替换基因修改.
  • 设计融合蛋白来提高编辑效率和特异性.

主要方法:

  • 开发使用nCas9-DNA聚合酶融合蛋白的仿真寡核酸定向编辑 (CODE) 系统.
  • 工程 CODEMax 和 CODEMax ((exo+) 编辑器,其中包含一个 Bst DNA 聚合酶衍生物.
  • 在人类细胞中评估编辑效率和意外编辑.

主要成果:

  • CODEMax和CODEMax ((exo+) 在人类细胞中证明了高效的基因组修改.
  • 这些CODE系统的无意编辑率很低.
  • 与PEMax相比,CODEs在各种位置上显示出较小插入,删除和替换的效率提高.

结论:

  • CODE系统为基因组操纵提供了一个强大的新工具.
  • 这些编辑器补充了现有的主要编辑技术.
  • CODE 能够在不引发双链断裂的情况下进行精确的编辑,扩大了基因组编辑工具箱.