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SiRA: 生細胞超解像度RNAイメージングのためのシリコンロダミン結合アプタマー
Regina Wirth1, Peng Gao2,3, G Ulrich Nienhaus2,3,4,5
1Institute of Pharmacy and Molecular Biotechnology (IPMB) , Heidelberg University , 69120 Heidelberg , Germany.
Journal of the American Chemical Society
|April 16, 2019
まとめ
研究者は,シリコンロダミン (SiR) 染料を結合する新しいRNAアプタマー (SiRA) を開発し,生細胞RNAイメージングのために明るい,光安定の遠赤色光放射を可能にしました. このシステムは,超高解像度顕微鏡を用いて,細菌と生細胞のRNAの視覚化を強化します.
科学分野:
- 分子生物学
- 生物化学
- バイオ物理学
背景:
- 遺伝的にコードされたライトアップRNAアプタマーは,生細胞RNAの可視化に価値があります.
- 遠赤と近赤外線 (NIR) を発するアプタマー/リガンドのペアは稀です.
- シリコンロダミン (SiRs) は,光安定性があり,細胞透過性が高いNIR放射性光体である.
研究 の 目的:
- 遠赤/NIR光成像のための新しいライトアップRNAアプタマーを開発する.
- シリコンロダミン染料を使って 汎用的なRNAラベリングツールを作りました
- 生細胞RNA画像と超解像度顕微鏡におけるこのシステムの応用を実証する.
主な方法:
- 組み合わせ図書室からRNAアプタマーのインビトロ選択
- 最小のアプタマー (SiRA) を特定するための生物情報分析,断片化,および変異研究.
- STED顕微鏡を含む光物理的特徴と生細胞画像実験.
主要な成果:
- 50ヌクレオチド最小のアプタマー,SiRAは,SiRsに対するナノモラー親和性を同定した.
- SiRAは高い光安定性,0.98の量子収量,そして86,000M−1cm−1の絶滅係数を示している.
- STED超解像度イメージングを含むSiRAを用いたバクテリアのRNAと生細胞のmRNAの視覚化に成功しました.
結論:
- SiRA-SiRシステムは,これまでに報告された最も明るい遠赤色ライトアップアプタマーです.
- この研究は,アプタマーベースのRNAイメージングのためにSiR染料と分子内スパイロサイクライゼーションを初めて適用したものです.
- SiRAシステムは,新しいRNAラベリング技術を通じて,生物学的研究を進めるための大きな可能性を秘めています.
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