巨大ファグからのCRISPR-CasΦは,超コンパクトなゲノムエディターです

Patrick Pausch^1,2, Basem Al-Shayeb^1,3, Ezra Bisom-Rapp⁴

¹Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.

Science (New York, N.Y.)

|July 18, 2020

まとめ

微小のCRISPR-CasシステムであるCasΦを発見しましたこのコンパクトな遺伝子編集ツールは既存の技術よりも強化されたDNAターゲティングと配信の利点を提供します.

科学分野:

微生物学
分子生物学
遺伝学

背景:

CRISPR-Casシステムは,外来核酸に対するプロカリオットの適応免疫を提供します.
これらのシステムはウイルスやプラズミドに対する防御に不可欠です

研究の目的:

巨大バクテリオファージから生まれた CRISPR-Casのシステムです
CasΦタンパク質とその関連CRISPR配列の機能と能力を特徴づける.

主な方法:

バクテリオファージのゲノムを生物情報学的に分析して,CasΦ系を特定する.
CasΦの酵素活性 (crRNA処理とDNA分裂) を評価する in vitro生化学測定法
ゲノム編集とDNA検出の可能性を評価するために,ヒトと植物細胞での機能的検証.

主要な成果:

単一のタンパク質,CasΦとCRISPR配列からなる超コンパクトのCRISPR-Casシステムの識別.
CasΦは,単一の活性部位を使用して,crRNA処理とcrRNA誘導DNA切断の両方を実行する二重活性を示します.
このシステムは,in vitroおよび真核細胞 (ヒトおよび植物) で活性を示した.
CasΦは他のCRISPR- Casタンパク質と比較して標的認識能力が向上した.
CasΦの小さなサイズ (Cas9とCas12aの分子量の半分) は,細胞配送を容易にする.

結論:

CasΦは独特の生化学特性を持つ新しく最小限のCRISPR-Casシステムを表しています
コンパクトな性質と二重機能により,高度なゲノム編集アプリケーションの有望なツールになります.
CasΦはCRISPRのツールボックスを拡張し,ゲノム編集とDNA検出における細胞伝達と潜在的により広範な標的認識の利点を提供します.

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