哺乳類の細胞と動物のRNA機能を調節する小分子アプタマー
Keisuke Fukunaga1, V Dhamodharan1, Nao Miyahira1
1Nucleic Acid Chemistry and Engineering Unit, Okinawa Institute of Science and Technology Graduate University, Onna, Okinawa 904 0495, Japan.
Journal of the American Chemical Society
|March 30, 2023
まとめ
研究者らは,新種のRNAアプタマーを用いて哺乳類の細胞における遺伝子調節のための合成リボスイッチを開発した. これらのリボスイッチは,低濃度でASP7967のような小分子に反応し,細胞培養物および体内での遺伝子発現を正確に制御することができます.
科学分野:
- 合成生物学
- 分子生物学
- バイオテクノロジー
背景:
- 合成リボスイッチはバイオテクノロジーと医学に潜力を秘めているが,哺乳類の細胞で機能する小さな分子-アプタマーペアによって制限されている.
- 既存のシステムでは,小分子濃度が高いため,実用的な応用が困難です.
研究 の 目的:
- 哺乳類の細胞における小分子媒介遺伝子調節のための新しいRNAアプタマーとリボスイッチを開発する.
- 低分子濃度で遺伝子調節を実現する.
- 開発されたリボスイッチの in vivo 応用を実証する.
主な方法:
- 小分子ASP7967に対するRNAアプタマーのインビトロ選択
- HEK293細胞におけるアパ酵素ベースのリボスイッチの設計と試験.
- マウスにおけるアデノ関連ウイルス (AAV8) のベクトルを用いてヒトのエリトロポエチン発現を調節するインビヴォ研究.
- アパツイムとエクソンスキップリボスイッチの組み合わせで 調節が強化される.
主要な成果:
- 機能性RNAアプタマー (AC17-4) を選択し,アプタ酵素ベースのリボスイッチを作成するために使用した.
- リボスイッチはASP2905またはASP7967で遺伝子発現を活性化しました.
- ヒトのエリトポエチン発現のインビオ調節は,マウスでASP7967の経口投与によって達成された.
- 結合したアパツイムとエクソンスキッピングリボスイッチシステムは,低ベース表現で約300のオン/オフ比率を達成した.
結論:
- 哺乳類の細胞で機能する新しい合成リボスイッチは,アプタマー選択を用いて開発された.
- この新しいシステムは,小分子の濃度が著しく低く,遺伝子調節を可能にします.
- 生体内での有効性と高性能の遺伝子制御の可能性が実証され,このシステムの生物医学とバイオテクノロジーの有望さを強調しています.
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