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関連する概念動画

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.
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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.
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Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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In 1928, a German botanist Emil Heitz observed the moss nuclei with a DNA binding dye. He observed that while some chromatin regions decondense and spread out in the interphase nucleus, others do not. He termed them euchromatin and heterochromatin, respectively. He proposed that the heterochromatin regions reflect a functionally inactive state of the genome. It was later confirmed that heterochromatin is transcriptionally repressed, and euchromatin is transcriptionally active chromatin.
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Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form...
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トポロジカルに制限されたDNAのスイッチ可能なトランスクリプションのプログラミング

Kai Jiao1, Bing Zhu1, Linjie Guo1

  • 1Division of Physical Biology, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 201800, China.

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まとめ

研究者は制御された遺伝子転写のために トポロジカルにオーダーされたDNA (TO-DNA) ナノ構造を作り出した. このDNA工学は,細胞イメージングのような応用のために,細菌の切り替え可能な遺伝子発現を可能にします.

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科学分野:

  • 合成生物学
  • 分子生物学
  • 生物化学

背景:

  • DNAトポロジとクロマチンの凝縮は細胞内の遺伝子発現を制御する.
  • 試験管内転写と遺伝子転移のためのDNAトポロジを制御することは依然として困難です.

研究 の 目的:

  • トポロジカルにオーダーされたDNA (TO-DNA) のナノ構造を制御された遺伝子発現のために構築する.
  • 転写活動に対するDNAトポロジーとプロモーター構成の影響を調査する.
  • 遺伝子工学や合成生物学の応用のための切り替え可能な転写システムを開発する.

主な方法:

  • T7プロモーター配列を備えた線形DNAテンプレートを作成するためにDNAセルフアセンブリが使用されました.
  • TO-DNAの転写活動は,プロモーターの位置と整合性に基づいて分析された.
  • ブール論理に基づく転写制御は,DNAの鍵鎖を用いて達成された.
  • バイオオルトホーナル・スイッチブル・トランスクリプションは,TO-DNAで複数の遺伝子で実装されました.
  • TO-DNAは,生きた細菌で光RNAアプタマーの切り替え可能な転写に使用された.

主要な成果:

  • TO-DNA構造は,T7プロモーター構成に敏感な転写活動を維持する.
  • T7プロモーターの特定の位置付けと無傷性は,ダイナミックでブール論理制御された転写活性化/抑制を可能にします.
  • 複数の遺伝子をTO-DNAに挿入して,バイオオルトゴナルで切り替え可能な転写を行うことができる.
  • バクテリアへの導入は,細胞画像化のための光RNAアプタマーの切り替え可能な転写をもたらした.

結論:

  • トポロジカルにオーダーされたDNAナノ構造は,形に依存した遺伝子配達と制御された遺伝子発現のための新しいプラットフォームを提供します.
  • このアプローチは遺伝子工学と合成生物学のツールボックスを強化し,転写を正確に制御することができます.
  • TO-DNAを用いた切り替え可能な転写は,生細胞画像と他のバイオテクノロジーの分野で潜在的な応用がある.