in vivoのタンパク質-DNA相互作用の全ゲノムマッピング
David S Johnson1, Ali Mortazavi, Richard M Myers
1Department of Genetics, Stanford University School of Medicine, Stanford, CA, 94305-5120, USA.
まとめ
ChIP-Seqを使ってタンパク質-DNAの相互作用をマッピングし,1946年にニューロン制限サイレンサー因子 (NRSF) の結合部位を特定しました. これは新しい結合モチーフと潜在的な遺伝子標的を明らかにし,遺伝子調節に関する理解を深める.
科学分野:
- ゲノミクスゲノミクスとは
- 分子生物学は分子生物学である.
- エピジェネティクス エピジェネティクス
背景:
- タンパク質とDNAの相互作用により,遺伝子調節ネットワークが形成されます.
- これらの相互作用をマッピングすることは,遺伝子発現を理解するために非常に重要です.
- ニューロン制限サイレンサー因子 (NRSF) は,遺伝子調節において重要な役割を果たします.
研究 の 目的:
- 生物体内のタンパク質-DNA相互作用の包括的なマッピングのために,大規模なクロマチン免疫降水測定法 (ChIP-Seq) を開発する.
- NRSFの結合部位を全ヒトゲノムにわたってマッピングする.
- 新しいNRSF結合モチーフと候補遺伝子ターゲットを特定する.
主な方法:
- 超高通量DNA配列解析を用いた大規模クロマチンの免疫降水測定法 (ChIP-Seq) を開発し,適用した.
- ヒトゲノムの1946の位置にNRSFのインビボ結合をマッピングした.
- 解像度,モチーフ識別,感度,特異性,統計的信頼性のための拘束力のあるデータを分析した.
主要な成果:
- ヒトゲノムの1946のNRSF結合部位を高解像度 (+/-50 bp) でマッピングしました.
- 特定されたカノニカルおよび新しい非カノニカルNRSFの拘束モチーフ.
- ChIP-Seqデータは,高い感度と特異性 (ROC領域>/=0.96) を示し,統計的信頼性 (P <10(-4) を示した.
結論:
- ChIP-Seqは,タンパク質-DNA相互作用の包括的な全ゲノムマッピングのための強力なツールです.
- 特定されたNRSF結合部位とモチーフは,遺伝子調節ネットワークの洞察を提供します.
- この研究は,臓の小島細胞の発達に関与するものを含む,新しい候補相互作用の推論を容易にする.
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