哺乳類のシス調節要素の核運動の転写結合変化
Bo Gu1, Tomek Swigut1, Andrew Spencley1,2
1Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Science (New York, N.Y.)
|January 27, 2018
まとめ
マルチプレックスガイドRNA配送のためのgRNAオリゴヌクレオチド (CARGO) のキメリック配列を開発しました. この方法は,生細胞での転写活性化によってシス調節要素の移動性が増加することを明らかにします.
科学分野:
- 分子生物学
- 遺伝学
- 細胞生物学
背景:
- マルチプレックスされたガイドRNA (gRNA) の配送は,複雑な遺伝子工学にとって極めて重要です.
- 細胞の分化などの過程におけるシス調節要素のダイナミクスを理解することは不可欠です.
研究 の 目的:
- 単一の細胞に複数の異なるgRNAを供給する新しい方法を開発する.
- 生きた胚性幹細胞における増強剤と促進剤の動態を定量的に測定する.
- シス調節要素の移動性と転写活動の関係を調査する.
主な方法:
- マルチプレックスされたgRNA配送のためのgRNAオリゴヌクレオチド (CARGO) のキメリック配列戦略の開発.
- CARGOと触媒死Cas9 (dCas9) のイメージングを組み合わせて,生細胞のゲノムロキーを追跡する.
- 微分化中の増強剤とプロモーターの動きと,波動に対する反応の定量分析.
主要な成果:
- CARGOは,何十種類もの異なるgRNAを単一の細胞に効率的に送ることができます.
- 検査されたすべての機能的要素 (強化剤,促進剤) はサブ拡散的行動を示した.
- トランスクリプションの活性化は,シス調節要素の移動性の増加と相関する.
- RNAポリメラーゼIIの活性障害は,活性に関連したロシモビリティの増加を逆転させた.
結論:
- CARGOは複数のgRNAの配送と遺伝子調節の研究に有効なツールです.
- シス調節要素の移動性は,転写活動と密接に関連しています.
- 生細胞画像は,異なる細胞状態における規制要素の機能に関するダイナミックな洞察を提供します.
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