関連する実験動画
Updated: May 8, 2026

11:03
Massively Parallel Reporter Assays in Cultured Mammalian Cells
Published on: August 17, 2014
クロマチンの位置効果は,並列に統合された数千人の記者が分析したものです
Waseem Akhtar1, Johann de Jong, Alexey V Pindyurin
1Division of Molecular Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, the Netherlands.
Cell
|August 20, 2013
まとめ
この研究では,遺伝子発現を測定するための高通量レポーターアッセイを導入しています. 測定は,ラミナ関連ドメインのようなクロマチンの構造が,遺伝子調節と強化剤の活性にどのように影響するかを明らかにします.
科学分野:
- 分子生物学は分子生物学である.
- ゲノミクスゲノミクスとは
- エピジェネティクス エピジェネティクス
背景:
- レポーター遺伝子アッセイは,遺伝子発現を研究する上で極めて重要です.
- 現在のレポーターアッセイは,スループットが限られており,大規模な分析を妨げています.
- ゲノム位置とクロマチンの文脈が遺伝子発現に与える影響を理解することは不可欠です.
研究 の 目的:
- 転写活動の並行モニタリングのための多重化,高通量レポーターアッセイを開発する.
- 遺伝子発現に対するローカルクロマチンの文脈と規制要素の影響を調査する.
- 遺伝子調節に関する新しい洞察を得るために,リポーター統合の大規模なデータセットを分析する.
主な方法:
- 何千ものレポーター統合の並行モニタリングのためのマルチプレキシングアプローチの開発.
- 27,000以上の異なるレポーター構造をマウスの胚性幹細胞に統合した.
- ゲノム位置とクロマチンの特徴に関連したレポーター発現レベルの分析.
主要な成果:
- 何千ものリポーター・インテグレーションで,表現レベルの約1,000倍の変動を示した.
- トランスクリプションの弱体化剤として,ラミナ関連ドメインを特定した.
- 示されたクロマチンの圧縮は,レポーター活動と推定強化剤の影響を約20kbで予測します.
結論:
- マルチプレックスレポーターアッセイは,遺伝子調節の高通量分析を可能にします.
- クロマチンの構造は,遺伝子発現レベルに大きな影響を与えます.
- このアッセイは,多様な遺伝子調節研究のための柔軟なプラットフォームを提供します.
関連する概念動画
Chromatin Position Affects Gene Expression
Chromatin is the massive complex of DNA and proteins packaged inside the nucleus. The complexity of chromatin folding and how it is packaged inside the nucleus greatly influences access to genetic information. Generally, the nucleus' periphery is considered transcriptionally repressive, while the cell's interior is considered a transcriptionally active area.
Topologically Associated Domains (TADs)
The 3-dimensional positioning of chromatin in the nucleus influences the timing and level of...
Topologically Associated Domains (TADs)
The 3-dimensional positioning of chromatin in the nucleus influences the timing and level of...
Position-effect Variegation
In 1928, a German botanist Emil Heitz observed the moss nuclei with a DNA binding dye. He observed that while some chromatin regions decondense and spread out in the interphase nucleus, others do not. He termed them euchromatin and heterochromatin, respectively. He proposed that the heterochromatin regions reflect a functionally inactive state of the genome. It was later confirmed that heterochromatin is transcriptionally repressed, and euchromatin is transcriptionally active chromatin.
Chromatin Immunoprecipitation- ChIP
Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
Spreading of Chromatin Modifications
The histone proteins in the nucleosomes are post-translationally modified (PTM) to increase or decrease access to DNA. The commonly observed PTMs are methylation, acetylation, phosphorylation, and ubiquitination of lysine amino acids in the histone H3 tail region. These histone modifications have specific meaning for the cell. Hence, they are called "histone code". The protein complex involved in histone modification is termed as "reader-writer" complex.
Writers
The writer is an enzyme that can...
Writers
The writer is an enzyme that can...
Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...
Chromatin Modification in iPS Cells
Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...

