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CRISPRベースのエピゲノム編集による全ゲノムプログラム可能な転写記憶

James K Nuñez¹, Jin Chen¹, Greg C Pommier²

¹Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158, USA; Howard Hughes Medical Institute, University of California, San Francisco, CA 94158, USA.

|April 10, 2021

まとめ

CRISPRoffは新しいツールでプログラム可能な表遺伝子記憶を使って遺伝子を安定的にオフにしますこの方法はCpG島の外でも機能し,遺伝子編集と表遺伝学の研究に幅広い応用が可能です.

科学分野:

分子生物学
エピジェネティクス
遺伝子規制

背景:

遺伝的遺伝子発現の変化は生物学的発見と治療に不可欠です
既存のエピジェネティック編集ツールには特殊性と遺伝性の限界があります

研究の目的:

安定した遺伝子サイレンスのためのプログラム可能なエピジェネティック・メモリーライターを開発する
全ゲノムスクリーニングとクロマチンの分析を用いて遺伝性遺伝子サイレンスに関する規則を確立する.

主な方法:

死んだCas9融合タンパク質 (CRISPRoff) を利用してDNAメチル化と抑制ヒストンの修正を確立した.
遺伝子の抑制のために一時的な CRISPR オフ表現を使用した.
ゲノム全体のスクリーニングとクロマチンのマーク分析を組み合わせたCRISPRオフ

主要な成果:

CRISPRoffは特定のDNAメチル化と遺伝子抑制を成功裏に開始しました
遺伝子の静止は細胞分裂と分化 (幹細胞からニューロンへ) を通じて維持された.
CpG島 (CGI) を欠いたものを含むほとんどの遺伝子を静止させる単一ガイドRNA (sgRNA) を特定し,CGIを超えた幅広いターゲットウィンドウを示した.

結論:

CRISPRoffは遺伝性遺伝子サイレンスのための多用途のプラットフォームを提供し,メチル化ベースのサイレンスに関する理解を広げています.
この技術は,全ゲノムスクリーン,複合細胞工学,増強器静音化などの多様なアプリケーションを可能にします.

キーワード:

CRISPRについて DNAメチル化細胞療法 dCas9 についてエピジェネティクス

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