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Intron seqFISHによる新生トランスクリプトームのダイナミクスと空間ゲノミクス
Sheel Shah1, Yodai Takei2, Wen Zhou2
1Division of Biology and Biological Engineering, Caltech, Pasadena, CA 91125, USA; UCLA-Caltech Medical Scientist Training Program, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Cell
|June 12, 2018
まとめ
光インシトゥハイブリデーション (seqFISH) によるイントロン配列化は,単細胞における新生RNA転写を視覚化します. この方法は,マウス細胞のダイナミックな核組織と非同期的な遺伝子発現の振動を明らかにします.
科学分野:
- 分子生物学
- ゲノミクス
- 細胞生物学
背景:
- 細胞内のトランスクリプトームと核の組織を視覚化することは依然として大きな課題です.
- 現存する方法は,現場で活性化した転写部位のダイナミクスを捉えるのに苦労しています.
研究 の 目的:
- 新生トランスクリプトームの多重化,単分子可視化のための新しい方法を開発する.
- 単細胞内の活性転写部位の空間的組織とダイナミクスを調査する.
- 細胞の種類と状態を新生トランスクリプトームプロファイリングで特定する.
主な方法:
- 光 in situ ハイブリデーション (intron seqFISH) によるイントロンの配列化を開発し,単一分子の多重化 in situ 方法である.
- 新生トランスクリプションの活性部位で 10,421 の遺伝子のイメージングを可能にしました.
- 総合的な分析のためにmRNAとlncRNAのseqFISHと免疫光と組み合わせる.
主要な成果:
- マウスの胚性幹細胞と線維芽細胞で異なる細胞タイプと状態を成功裏に特定した.
- 新生RNA合成部位は,通常,細胞間の高変性を持つ染色体領域の表面に位置することを示した.
- 2つの細胞タイプで2時間の周期でグローバル新生転写における非同期的な振動を発見した.
結論:
- Intron seqFISHは,単細胞の新生トランスクリプトームの空間のゲノミクスを提供しています.
- 核組織の基本原理と 素早い転写ダイナミクスを明らかにします
- 細胞の異質性や遺伝子調節を分析する強力なツールです
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