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細胞遺伝スクリーンによる遺伝子規制のマッピングのための全ゲノムフレームワーク
Molly Gasperini1, Andrew J Hill1, José L McFaline-Figueroa1
1Department of Genome Sciences, University of Washington, Seattle, WA 98105, USA.
Cell
|January 8, 2019
まとめ
科学者たちは新しいCRISPRベースのシステムを用いて 何千もの遺伝子調節要素をマッピングしました この新しい方法は 増強物質と標的遺伝子を結び付け 人間のゲノムの複雑な 規制構造の理解を深めています
科学分野:
- ゲノミクス
- 分子生物学
- エピジェネティクス
背景:
- ヒトゲノムにおける100万以上の候補の規制要素には 検証と明確な標的遺伝子の識別が欠けている.
- 現在のヒト遺伝学のアプローチは,一般的な変種とリンク不均衡によって制限され,規制要素分析の範囲と解決を制限しています.
研究 の 目的:
- ヒトゲノム全体のエンハンサー遺伝子ペアをマッピングするための高通量フレームワークを開発し,検証する.
- 機能的な規制要素とそのターゲットを特定するための既存の方法の限界を克服する.
主な方法:
- CRISPR/Cas9スクリーニングフレームワークにインスパイアされたマルチプレックス,表現量的な特性の場所 (eQTL) が使用されました.
- dCas9- KRABは,複数の細胞に 5, 920の候補強化剤を乱すのに使用されました.
- 単細胞RNAシーケンシング (RNA-seq) は,254,974個の単細胞トランスクリプトームで perturbation効果を測定するために実施された.
主要な成果:
- 664のシス増強遺伝子ペアが特定され,そのうちの470は高い信頼度であると考えられています.
- 特定されたペアは,特定の転写因子と非家事遺伝子状態の強化を示した.
- 結果は,検証された強化剤-遺伝子ペア間のゲノムと3D構造の接近を示した.
結論:
- 開発された枠組みは,増強剤-遺伝子規制相互作用の大規模マッピングを可能にします.
- このアプローチは,ヒトゲノムのシス調節の特徴を大幅に進める.
- この発見は,遺伝子調節と生物学的過程におけるその役割に関する将来の研究のための基盤を提供します.
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