SaCas9の分子メカニズムを拡大したDNAターゲティング範囲で探求するための組み合わせた計算実験アプローチ
Binquan Luan1, Guangxue Xu2, Mei Feng3
1Computational Biological Center , IBM Thomas J. Watson Research , Yorktown Heights , New York 10598 , United States.
Journal of the American Chemical Society
|March 30, 2019
まとめ
研究者は,Staphylococcus aureus Cas9 (SaCas9) 酵素を研究することで,CRISPR- Cas9技術を研究した. 彼らはSaCas9の変異の分子メカニズムを明らかにし,活性化とより広範な標的能力を持つ新しい変異体の設計につながりました.
科学分野:
- バイオテクノロジー
- 分子生物学
- ゲノミクス
背景:
- CRISPR技術は急速に進歩していますが その基本的な生物学的な性質については さらに研究が必要です
- エンジニアリングされたCRISPRツールには 改良された機能の 機械的な説明が しばしば欠けています
- Staphylococcus aureus Cas9 (SaCas9) は in vivo 適用に適しているが,長いプロトスペーサー付近モチーフ (PAM) の必要性がある.
研究 の 目的:
- SaCas9 PAM認識の分子ダイナミクスを調査する.
- SaCas9におけるKKH変異のメカニズムを解明する.
- 強化された特性を備えた新しい SaCas9 変種を設計し,検証する.
主な方法:
- 構造ベースの分子ダイナミクスシミュレーション
- 自由エネルギーによる混乱分析
- SaCas9変異体の標的型実験検証
主要な成果:
- SaCas9におけるPAM認識のダイナミックメカニズムを明らかにした.
- KKH変異の分子基盤を明らかにした.
- SaCas9-NRとSaCas9-RLの突然変異は,哺乳類の細胞における標的範囲を拡大し,活性性を高めるように設計され,検証されています.
結論:
- この研究は,SaCas9 PAM認識のダイナミックな理解を提供します.
- 開発されたワークフローは,新しい性質を持つCas9のバリエーションの合理的な設計を可能にします.
- 新しい SaCas9 変種は,遺伝子編集アプリケーションのための改善されたツールを提供します.
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