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結合した単細胞CRISPRスクリーニングとエピジェノミックプロファイリングは,因果的な遺伝子規制ネットワークを明らかにする
Adam J Rubin1, Kevin R Parker2, Ansuman T Satpathy3
1Program in Epithelial Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Cell
|December 25, 2018
まとめ
Perturb-ATACは,CRISPR遺伝子編集とクロマチンのアクセシビリティプロフィールを組み合わせて,遺伝子調節をマッピングする新しい単細胞法です. このアプローチは,転写因子がヒトの細胞のアイデンティティと発達をどのように制御するかを明らかにします.
科学分野:
- ゲノミクス
- 分子生物学
- 細胞生物学
背景:
- 遺伝子制御ネットワークを理解することは 細胞の発達や病気の解読に不可欠です
- 現在の方法は,単一の細胞内の複雑な相互作用を解剖する解像度が不足しています.
研究 の 目的:
- 遺伝子規制ネットワークのプロファイリングのための新しい単細胞技術であるPerturb-ATACを導入します.
- 細胞プロセスにおける転写因子,クロマチンの変形因子,および非コーディングRNAの役割を調査する.
主な方法:
- Perturb-ATACは複数のCRISPR干渉/ノックアウトと単細胞ATAC-seqを組み合わせている.
- ガイドRNAとクロマチンの開いた部位を同時に検出することで,ゲノタイプ-フェノタイプマッピングが可能になります.
- 63以上の遺伝子型-フェノタイプ関係を分析し,約4,300個の単細胞に適用した.
主要な成果:
- 人間のBリンパ球では,クロマチンのアクセシビリティ,TF占有率,および核細胞の位置づけのレギュレータが特定されました.
- B細胞の状態,変異,および疾患に関連する要素を支配するTF階層を発見した.
- 皮質細胞では,ケラチノシートの運命を決定するシーケンス的なシス要素モジュールを明らかにした.
- ゲノム共同局所化に基づいたTFエピスタティック関係とシネジスティック相互作用を発見した.
結論:
- Perturb-ATACは 発達と疾患における 遺伝子制御ネットワークを 分解する強力な戦略です
- この方法は 細胞の意思決定プロセスに 前例のない洞察力をもたらします
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