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In Vivo CRISPR/Cas9 Screening to Simultaneously Evaluate Gene Function in Mouse Skin and Oral Cavity07:52

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Here we describe a rapid and direct in vivo CRISPR/Cas9 screening methodology using ultrasound-guided in utero embryonic lentiviral injections to simultaneously assess functions of several genes in the skin and oral cavity of immunocompetent...
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In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression08:54

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This protocol outlines the steps needed to generate a model system in which the transcription of an endogenous gene of interest can be conditionally controlled in live animals or cells using enhanced lac repressor and/or tet activator...
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This protocol describes the steps for cloning multiple single guide RNAs into one guide RNA concatemer vector, which is of particular use in creating multi-gene knockouts using CRISPR/Cas9 technology. The generation of double knockouts in intestinal organoids is shown as a possible application of this...
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The microinjection of mouse oocytes is commonly used for both classic transgenesis (i.e., the random integration of transgenes) and CRISPR-mediated gene targeting. This protocol reviews the latest developments in microinjection, with a particular emphasis on quality control and genotyping...
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DNA Vector-based RNA Interference to Study Gene Function in Cancer

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RNA interference (RNAi) possesses many advantages over gene knockout and has been broadly used as a tool in gene functional studies. The invention of DNA vector-based RNAi technology has made long term and inducible gene knockdown possible, and also increased the feasibility of gene silencing in...
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Rapid Characterization of Genetic Parts with Cell-Free Systems
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合成遺伝子ポリマーからの触媒.

Alexander I Taylor1, Vitor B Pinheiro1, Matthew J Smola2

  • 1MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH, UK.

Nature
|December 4, 2014
PubMed
まとめ

合成遺伝子ポリマー,またはXNAは,XNA酵素と呼ばれる触媒を形成することができます. この発見は,生命の可能性を拡大する.

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科学分野:

  • 生命の起源の研究 生命の起源の研究
  • 合成生物学 合成生物学とは
  • バイオケミストリー バイオケミストリー

背景:

  • RNAのような初期の遺伝子ポリマーにおける触媒は,タンパク質酵素に先立ち,生命の起源にとって極めて重要でした.
  • DNAは触媒構造に折りたたむことができるが,特権的な支架としての天然バイオポリマーの可能性は不明である.
  • 新しい骨格を持つ合成遺伝子ポリマー (XNA) は,リガンドを折り畳んで結合することができ,触媒的可能性を示唆しています.

研究 の 目的:

  • 合成遺伝子ポリマー (XNAs) が機能的触媒 (XNAzymes) を形成できるかどうかを調査する.
  • 様々なXNA化学系におけるエンドヌクレアゼおよびリガゼ活動を持つXNA酵素を発見する.
  • 完全合成のXNAシステム内で触媒を確立する.

主な方法:

  • アラビノ核酸 (ANA),2'-フッ素アラビノ核酸 (FANA),ヘキソール核酸 (HNA),サイクロヘクセン核酸 (CeNA) の4つの異なる化学成分からランダムなプールからXNA酵素の選択.
  • 発見されたXNA酵素のトランスRNAエンドヌクレアゼとリガゼの活性を示す.
  • 完全合成触媒システムのためのFANAベースのXNA-XNAリガース金属酵素のエンジニアリング.

主要な成果:

  • 4つの異なるXNA化学でRNAエンドヌクレアゼとリガゼ活動を示すXNA酵素の発見.
  • FANAオリゴーマーを結合し,活性RNAエンドヌクレアースFANAzymeを合成するFANAベースのメタロ酵素の作成.
  • 自然生体ポリマーから独立した触媒の実証.

結論:

  • 触媒は,RNAやDNAのような天然のバイオポリマーを超えて,合成遺伝子ポリマーで発生することができます.
  • XNA酵素は,合成生物学における潜在的な応用を持つ新しい種の触媒を代表しています.
  • この研究は,地球上の生命の起源に関する化学的可能性を拡大し,他の場所でも可能性が高い.