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バクテリアのシチジンデアミナーゼ毒素は,CRISPRフリーでミトコンドリアの塩基編集を可能にします

Beverly Y Mok^1,2,3, Marcos H de Moraes⁴, Jun Zeng⁴

¹Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

|July 10, 2020

まとめ

研究者は細菌毒素から新しいRNAフリーサイトシンベースエディター (DdCBE) を開発した. このツールはミトコンドリアDNA (mtDNA) を疾患モデリングおよび潜在的な治療用途のために正確に修正します.

科学分野:

生物化学
分子生物学
遺伝学

背景:

細菌の毒素は遺伝子編集を含む生物医学的な用途に多様な生化学的ツールを提供しています
既存のシチジンデアミナーゼは,塩基編集のために二重鎖DNA (dsDNA) を解き放つ必要があります.
ミトコンドリアDNA (mtDNA) の塩基編集は,配送の問題により,ゲノム破壊に操作を制限することが困難である.

研究の目的:

新種の細菌間毒素であるDddAを記述します
精密な mtDNA 操作のための RNA のない DddA 派生型サイトシンベースエディター (DdCBEs) を設計する.
mtDNA変異とその細胞上の結果のモデリングにおけるDdCBEsの適用を実証する.

主な方法:

標的のDNAに結合するまで機能しない
転写活性化器のようなエフェクター配列タンパク質とウラシルグリコシラーゼ阻害剤を融合させたDddA.
人間のmtDNAにおけるRNAフリーC•G-to-T•A塩基編集のためにDdCBEsを使用した.

主要な成果:

ヒトのmtDNAで高特異性と純度でC•G•T•A変換を実現するDdCBEsを開発しました.
DdCBEsを用いてヒトの細胞で病気に関連したmtDNA変異をモデル化しました.
mtDNA変異モデリングによる細胞呼吸と酸化リン酸化の変化を観察した.

結論:

CRISPR のないDdCBEsは,ニュクレアズベースの方法とは異なり,mtDNAの正確な操作を可能にします.
この技術は,ミトコンドリア障害の研究と潜在的治療に広範囲の影響を及ぼします.
DddA由来の塩基エディターはミトコンドリアゲノム工学の重要な進歩を表しています.

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