CRISPRを,配列特異的な遺伝子発現制御のためのRNA誘導プラットフォームとして再利用する
Lei S Qi1, Matthew H Larson, Luke A Gilbert
1UCSF Center for Systems and Synthetic Biology, University of California, San Francisco, San Francisco, CA 94158, USA. stanley.qi@ucsf.edu
Cell
|March 5, 2013
まとめ
私たちはCRISPR干渉 (CRISPRi) を開発し,遺伝子の発現を正確に制御するために,無効化されたCas9タンパク質を使用した新しい遺伝子編集ツールを開発しました. このシステムは細菌の標的遺伝子を効率的に抑制し,哺乳類の細胞での使用の可能性を示しています.
科学分野:
- 分子生物学は分子生物学である.
- 遺伝学 遺伝学とは
- バイオテクノロジー バイオテクノロジー
背景:
- ゲノム全体の遺伝子調節は,細胞機能を理解し,エンジニアリングするために重要です.
- CRISPRシステムは,精密なDNAターゲティングのための強力なツールを提供します.
研究 の 目的:
- CRISPR技術を用いた標的型遺伝子調節のための新しい方法を開発する.
- 遺伝子抑制のためのこのシステムの有効性と特異性を実証するために.
主な方法:
- 触媒的に死んだCas9 (dCas9) タンパク質をRNAによって誘導して,DNA認識複合体を作成しました.
- Escherichia coli のガイドRNA と dCas9 を共発し,転写過程に干渉する.
- 遺伝子抑制効率と非標的効果を評価した.
主要な成果:
- CRISPR干渉 (CRISPRi) は,標的外効果を検出することなく,E. coliにおける標的遺伝子発現を効率的に抑制しました.
- CRISPRiシステムは,複数の遺伝子を同時に抑制する能力を実証しました.
- このシステムの効果は,哺乳類の細胞における遺伝子抑制に逆転し,適応することが示された.
結論:
- CRISPRの干渉は,選択的遺伝子干渉のためのシンプルで効果的なRNA誘導のDNA認識プラットフォームを提供します.
- この技術は,研究およびエンジニアリングアプリケーションのための遺伝子発現に対するゲノム全体の制御を可能にします.
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