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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.0K
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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In-vitro Mutagenesis01:16

In-vitro Mutagenesis

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To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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DNA-only Transposons02:57

DNA-only Transposons

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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LTR Retrotransposons03:08

LTR Retrotransposons

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LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
The internal coding region of LTR retrotransposons and their mechanism of transposition closely resembles a...
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Exon Recombination02:32

Exon Recombination

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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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相关实验视频

Updated: Jun 28, 2025

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
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In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

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霍克斯基因特异性细胞向使用分裂整体特洛伊木马外子.

Fengqiu Diao1, Deeptha Vasudevan2, Ellie S Heckscher2

  • 1Laboratory of Molecular Biology, Section on Neural Function, National Institute of Mental Health, NIH, Bethesda, MD 20892.

Proceedings of the National Academy of Sciences of the United States of America
|April 11, 2024
PubMed
概括
此摘要是机器生成的。

一种新的Split Intein木马外子 (siTrojan) 方法通过防止有害突变来改善Drosophila Gal4驱动线. 这种技术确保了全长蛋白质的生产,使细胞类型分析的精确基因向成为可能.

关键词:
这种植物是Drosophila.细胞类型 细胞类型发展发展发展发展发展.遗传接入 遗传接入神经电路映射的神经电路映射

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HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries
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CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
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相关实验视频

Last Updated: Jun 28, 2025

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
08:54

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

Published on: March 29, 2019

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HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries
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CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
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科学领域:

  • 发展生物学 发展生物学
  • 遗传学 是一个遗传学.
  • 神经科学是一个神经科学.

背景情况:

  • 在Drosophila中使用的特洛伊木马外显子方法产生了基因特异的Gal4驱动线,用于细胞类型访问和功能丧失分析.
  • 然而,来自特洛伊木马外子的截断突变可能会导致主导负效应和发育致死性,特别是对于基本的转录因子.
  • 需要使用替代方法来创建Gal4线,以向转录因子而无有害突变.

研究的目的:

  • 引入一种修改后的特洛伊木马外型子方法,可以减轻突变原体的影响,同时保持准特异性.
  • 为霍克斯转录因子开发Gal4和Split Gal4线的工具包.
  • 为了证明神经电路映射中的新方法的实用性.

主要方法:

  • 开发了Split Intein特洛伊木马前体 (siTrojans),可以使用自剪接的分割整体.
  • siTrojans确保截断产品的转接,以恢复完整的原生蛋白质功能.
  • 为分段表达的霍克斯转录因子生成了Gal4和Split Gal4线.

主要成果:

  • siTrojan方法成功地减轻了原始Trojan外型子方法的突变效应.
  • 为霍克斯转录因子创建了Gal4和Split Gal4线的综合工具包.
  • 在神经电路映射中通过基于前后位置的准神经元来证明siTrojans的应用.

结论:

  • 分裂内因特洛伊木马外原体 (siTrojans) 提供了一种有价值的替代方案,用于在Drosophila中生成基因特异的Gal4驱动线.
  • 这种方法保持了准准确性和模块化性,同时防止了有害的突变.
  • siTrojan方法和霍克斯转录因子工具包为Drosophila的遗传研究提供了广泛的实用性.