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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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Viral Recombination00:57

Viral Recombination

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Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
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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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Recombinant DNA01:09

Recombinant DNA

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Overview
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相关实验视频

Updated: Jun 14, 2025

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
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Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

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使用复合子进行连续多重菌体基因组编辑.

Chloe B Fishman1, Kate D Crawford1,2, Santi Bhattarai-Kline1,3

  • 1Gladstone Institute of Data Science and Biotechnology, San Francisco, CA, USA.

Nature biotechnology
|September 5, 2024
PubMed
概括
此摘要是机器生成的。

我们开发了一种可扩展的方法,使用重组子进行高效的菌体基因组编辑,增强对细菌的菌体治疗. 这种技术可以简化修改,而无需费力的选,从而使改进的菌体的发展速度更快.

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The MultiBac Protein Complex Production Platform at the EMBL
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The MultiBac Protein Complex Production Platform at the EMBL

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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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相关实验视频

Last Updated: Jun 14, 2025

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

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The MultiBac Protein Complex Production Platform at the EMBL
13:51

The MultiBac Protein Complex Production Platform at the EMBL

Published on: July 11, 2013

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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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科学领域:

  • 分子生物学分子生物学
  • 微生物学 微生物学
  • 合成生物学 合成生物学

背景情况:

  • 菌体基因组编辑对于改善菌体治疗对病原性细菌的疗效至关重要.
  • 目前的菌体基因组编辑方法效率低下,需要广泛的选和反选择.
  • 需要可扩展和高效的方法来进行菌体基因组修改.

研究的目的:

  • 开发一种可扩展和高效的细菌基因组编辑方法.
  • 为了克服当前繁忙的菌体基因组编辑技术的局限性.
  • 为了实现治疗应用的修饰菌体的快速生成.

主要方法:

  • 利用修改后的细菌反子 (重组子) 来产生重组的供体DNA.
  • 采用单链结合和回火蛋白来有效地将DNA整合到菌体基因组中.
  • 证明了该系统能够进行连续的,可多重复的基因组修改的能力.

主要成果:

  • 在无对抗选择的情况下,在多个菌体中实现了高效的基因组修改.
  • 在与反选择相结合时,成功执行了大插入和删除>99%的效率.
  • 在几个小时内,在单个兰巴菌体基因组上安装了多达五种不同的突变.
  • 在T7 gp17尾部纤维中确定了残留水平的表皮性相互作用.

结论:

  • 这种基于重组子的系统为菌体基因组编辑提供了可扩展和高效的方法.
  • 这种方法显著减少了菌素修饰所需的劳动和时间.
  • 开发的系统促进了菌体的快速工程,以提高治疗潜力.