検出可能な全ゲノムオフターゲットの変異なしの in vivo CRISPR 編集
Pinar Akcakaya1, Maggie L Bobbin2,3,4, Jimmy A Guo2,3
1Discovery Biology, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Gothenburg, Sweden.
Nature
|September 14, 2018
まとめ
新しい方法である in vivo off-targets (VIVO) の検証は,信頼性の高い CRISPR-Cas off-target 変異を in vivo で検出します. VIVOは遺伝子編集が マウスの肝臓で安全で効果的であることを示しています
科学分野:
- バイオテクノロジー
- 遺伝学
- 分子生物学
背景:
- CRISPR-Casのゲノム編集は治療的な可能性を秘めているが,臨床安全性のために標的外変異の特定が必要である.
- 現在,標的外変異をin vivoで確実に検出するための検証された方法が欠如しており,臨床翻訳を阻害しています.
研究 の 目的:
- CRISPR-Cas核酵素の全ゲノムにわたる非標的効果を in vivo で確実に特定するために,非常に敏感な戦略である in vivo off-target (VIVO) の検証を導入する.
- CRISPR- Cas核酵素によって誘発される標的外変異の頻度と影響を in vivo で評価する.
主な方法:
- 対象外生物 (VIVO) の検証戦略の策定と適用
- マウスの肝臓の変異を検知する VIVOの感受性をテストするために 意図的に乱れたガイドRNAを使用します
- 適切な設計のガイドRNAの安全性を評価するためにVIVOを使用する.
主要な成果:
- CRISPR-Cas核酸は,乱交型ガイドRNAを使用すると,マウスの肝臓に実質的なオフターゲットの変異を誘導することができます.
- 適切に設計されたガイドRNAは,検出可能な標的外変異のないマウス肝臓で効率的な in vivo 編集を可能にします.
- VIVOは,全ゲノム対象外 in vivo分析のための敏感で堅固な方法であることが証明されています.
結論:
- VIVO戦略は,全体的な生物における遺伝子編集核酸の非標的効果を定義し,定量化するための信頼できる方法を提供します.
- 安全で効果的な in vivo 遺伝子編集を実現するには,注意深いガイドRNA設計が不可欠です.
- VIVOは,in vivo遺伝子編集を利用したより安全な治療戦略の開発のための青写真を提供しています.
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