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相关概念视频

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.
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Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
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In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
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ribozyme-Enabled Tissue Specificity (RETS):一种在没有专门的促进器的情况下实现精确基因表达的系统.

Max M Combest1, Josh Conlin1, Vivia Van De Mark2

  • 1Department of Biology, Colorado State University, Fort Collins, Colorado 80523, United States.

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概括

一种名为Ribozyme-Enabled Tissue Specificity (RETS) 的新方法允许精确控制植物中的基因表达. 这种技术使组织特异性转基因表达能够在不需要已知的促进剂的情况下实现,有助于生物研究和作物工程.

关键词:
表达式生物传感器生物传感器植物发展 植物发展植物合成生物学 植物合成生物学ribozymes ribozymes ribozymes 是一种类型的有机化合物.组织特异性表达.

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科学领域:

  • 植物生物学 植物生物学
  • 分子生物学分子生物学
  • 合成生物学 合成生物学

背景情况:

  • 在植物中实现精确的组织特异性基因表达对于生物研究和工程至关重要,但经常受到神秘增强剂和长时间原型化时间限制的阻碍.
  • 控制转基因表达的现有方法可能是具有挑战性的,原因是促进体识别的困难和转基因谱系发展所需的时间.

研究的目的:

  • 开发一种新的策略,即 Ribozyme-Enabled Tissue Specificity (RETS),用于在不依赖于特征性促进体的情况下,在植物中实现可预测的,组织特异的转基因表达.
  • 为了证明RETS在各种植物科学应用中的灵活性,效率和适用性.

主要方法:

  • 开发了RETS,使用分裂的自我拼接 ribozyme (来自 *Tetrahymena thermophila* 的 I 组内核) 进行条件 mRNA 复制.
  • 设计的RETS利用转录组数据针对特定的内源mRNA,使条件转基因表达成为可能.
  • 优化了RETS设计,用于转基因/目标交换,增强表达和避免RNA干扰.

主要成果:

  • 通过使用RETS系统,成功地证明了Arabidopsis thaliana*的组织特异性和剂量依赖的转基因表达.
  • 展示了RETS在创建基因编码生物传感器中的实用性,用于植物中的基因表达空间时间模式分析.
  • 经过验证的RETS用于设计器官大小的特定组织变化,证明其具有精确表型修饰的潜力.

结论:

  • RETS提供了一种突破性的方法来研究本地基因表达模式的非破坏性方法,克服了当前方法的局限性.
  • 通过RETS对转基因表达的时空空间控制促进了植物表型的精密工程,使作物增强无需构成表达缺陷.