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CRISPR媒介型モジュラーRNA誘導によるエウカリウトにおける転写の調節

Luke A Gilbert¹, Matthew H Larson, Leonardo Morsut

¹Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA.

|July 16, 2013

まとめ

科学者たちは,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てCRISPR干渉 (CRISPRi) を開発しました. この強力なツールは,遺伝子研究のためにヒトおよび酵母細胞で標的の遺伝子の静止または活性化を可能にします.

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:

精密な遺伝子ターゲティングは,細胞遺伝学の研究と再プログラムに不可欠です.
触媒的に無活性なCas9 (dCas9) タンパク質は,RNA誘導のDNAターゲティングのためのプラットフォームを提供します.

研究の目的:

遺伝子調節のためのdCas9がエフェクタドメインと融合した有用性を実証するために.
転写抑制と活性化のための方法としてCRISPR干渉 (CRISPRi) を確立する.

主な方法:

dCas9の融合により,異なる規制効果ドメインが形成される.
ヒトおよび酵母細胞における標的型DNA配送のためのショートガイド (sg) RNAを使用する.
RNAシーケンシング (RNA-seq) を使用して遺伝子発現の変化を分析する.

主要な成果:

安定的かつ効率的な転写抑制と活性化は,dCas9とエフェクタドメインを融合させることで達成された.
CRISPRの干渉は,複数の内生遺伝子を強力に静止させました.
RNA-seqは,CRISPRi媒介による転写抑制の高特異性を確認しました.

結論:

CRISPRシステムは,タンパク質の採用のためのモジュラーDNA結合プラットフォームとして機能します.
CRISPRの干渉は,真核細胞における精密な遺伝子発現調節のための汎用的なツールです.
この技術は,遺伝子研究と治療応用において大きな可能性を秘めています.

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