CRISPR RNAと複合したCpf1の結晶構造

De Dong¹, Kuan Ren¹, Xiaolin Qiu¹

¹School of Life Science and Technology, Harbin Institute of Technology, Harbin 150080, China.

Nature

|April 21, 2016

まとめ

CRISPR-Cpf1システムはゲノム編集の新しいツールで DNAターゲティングのユニークな構造を示していますそのRNA認識メカニズムの理解はより正確な遺伝子編集技術を開発するのに役立ちます

科学分野:

分子生物学
遺伝学
生物化学

背景:

CRISPR-Casシステムは,プロカリオットの適応性免疫メカニズムである.
CRISPR-Cpf1 (Class 2) は,CRISPR-Cas9とは異なるヒトの細胞にDNA干渉をもたらす.
Cpf1とCas9はガイドRNAと基板特異性によって異なります.

研究の目的:

CRISPR RNA (crRNA) に結合したLachnospiraceae菌ND2006 Cpf1 (LbCpf1) の結晶構造を決定する.
LbCpf1によるcrRNA認識のメカニズムを解明する.
ゲノム編集アプリケーションのためのcrRNA誘導基板結合に関する洞察を提供します.

主な方法:

crRNAに結合したLbCpf1のX線結晶図 (2.38 Å解像度)
LbCpf1構造とcrRNA結合の構造分析
crRNA結合時の形状の変化を調査する.

主要な成果:

LbCpf1は中央の正電荷チャネルを持つ三角形構造を示している.
crRNAは,相互作用とマグネシウムイオンによって安定した,オリゴヌクレオチド結合領域内の歪んだ形状を採用します.
crRNA結合は,オリゴメリゼーションなしにLbCpf1の構造変化を誘導する.

結論:

この研究は,LbCpf1によるcrRNA認識の構造的基礎を明らかにしている.
crRNAによる基板結合に関する洞察は,LbCpf1エンジニアリングの基盤を提供します.
この研究は,効率と特異性を向上した LbCpf1 ベースの改良されたゲノム編集ツールの開発を容易にする.

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