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相关概念视频

Gene Conversion02:08

Gene Conversion

9.6K
Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
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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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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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Exon Recombination02:32

Exon Recombination

3.5K
The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
Exon shuffling follows “splice frame rules.” Each exon...
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Crossing Over01:30

Crossing Over

4.1K
Crossing over is the exchange of genetic information between homologous chromosomes during prophase I of meiosis I. Genetic recombination gives rise to allelic diversity in the newly formed daughter cells. In humans, crossing over produces genetically distinct haploid egg and sperm cells that undergo fertilization to produce unique offspring. Before cell division starts, the germ cell’s chromosome(s) undergo duplication in the S phase of the cell cycle. As the cells enter prophase I,...
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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: May 30, 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

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通过重复元素的工程重组来随机化人类基因组

Jonas Koeppel1, Raphael Ferreira2,3,4, Thomas Vanderstichele1

  • 1Wellcome Sanger Institute, Hinxton, UK.

Science (New York, N.Y.)
|January 30, 2025
PubMed
概括
此摘要是机器生成的。

研究人员使用CRISPR原始编辑来研究非编码DNA的基因组编码策略. 这种方法产生了大量的DNA重组,揭示了基因组组织如何影响基因表达和选择压力.

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Recombineering Homologous Recombination Constructs in Drosophila
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Recombineering Homologous Recombination Constructs in Drosophila

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Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus
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Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus

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相关实验视频

Last Updated: May 30, 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

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Recombineering Homologous Recombination Constructs in Drosophila
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Recombineering Homologous Recombination Constructs in Drosophila

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Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus
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Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus

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科学领域:

  • 基因组学和分子生物学
  • 表观遗传学和基因调控

背景情况:

  • 目前的工具不足以进行大规模的DNA编辑,阻碍了对99%的非编码人类基因组的研究.
  • 了解非编码DNA对于破译基因组组织和基因调节至关重要.

研究的目的:

  • 在非编码基因组中开发大规模DNA操纵的新工具.
  • 通过诱导的重组来研究基因组的可用性和组织原则.

主要方法:

  • 应用CRISPR原始编辑来插入重组处理器到重复序列中.
  • 诱导重组酶以产生随机的大基大小的DNA重组 (删除,反转,转位,循环DNA).
  • 随着时间的推移跟踪重新排列以评估选择压力和结果克隆的特征.

主要成果:

  • 每个细胞系成功产生了100多个兆基大小的重组.
  • 观察到的选择有利于避免必要基因的较短变体.
  • 显示删除影响变体内的基因表达而不是相邻基因的特征克隆.

结论:

  • 开发的基因组编码策略可以实现前所未有的大规模DNA编辑.
  • 这种方法有助于探索基因组组织,可用性以及结构变异对基因表达的影响.