ネイティブタンパク質の効率的な細胞内伝達
Diego S D'Astolfo1, Romina J Pagliero2, Anita Pras3
1KNAW-Hubrecht Institute, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands; University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the Netherlands.
Cell
|April 25, 2015
まとめ
オスモサイトーシスとプロパネベタイン (iTOP) による誘導伝導を開発し,細胞に直接タンパク質を供給する新しい方法を開発しました. このテクニックは,遺伝子編集のためのCas9を含むタンパク質を効率的に様々な細胞タイプに供給します.
科学分野:
- 細胞生物学 細胞生物学
- 分子生物学は分子生物学である.
- バイオテクノロジー バイオテクノロジー
背景:
- 細胞内タンパク質の直接供給は困難であり,細胞のプロセス干渉を制限する.
- 現在の方法は,DNAまたはmRNAによるエフェクタータンパク質による間接的な伝達に依存しています.
研究 の 目的:
- 細胞内タンパク質を直接かつ効率的に供給するための新しい方法を開発する.
- この方法が一時的な細胞操作と遺伝子編集における有用性を実証する.
主な方法:
- オスモサイトーシスとプロパネベタインによる誘導変換 (iTOP) は,高圧性誘導マクロピノサイトーシスとプロパネベタインを使用します.
- iTOPは,タンパク質を様々な原始細胞タイプに変換することを促進します.
主要な成果:
- iTOPは,さまざまな細胞タイプで高効率のタンパク質伝導を実現します.
- この方法は,非統合的遺伝子ターゲティングのための再結合Cas9タンパク質と短いガイドRNAを成功裏に提供しました.
結論:
- iTOPは,以前の制限を克服し,直接タンパク質を供給するための堅牢なプラットフォームを提供します.
- このテクニックは,効率的なCRISPR-Cas9遺伝子編集によって例示される,永久的な細胞変化のための一時的な細胞操作を可能にします.
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