相关实验视频
Updated: May 8, 2026

11:03
Massively Parallel Reporter Assays in Cultured Mammalian Cells
Published on: August 17, 2014
数以千计的记者并行集成测定了染色体位置效应
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
概括
这项研究引入了一种高通量记者测试来测量基因表达. 该试验揭示了染色质结构,如膜相关域,如何影响基因调节和增强剂活性.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
背景情况:
- 记者基因测试对于研究基因表达至关重要.
- 目前的记者分析具有有限的吞吐量,阻碍了大规模分析.
- 了解基因组位置和染色质背景对基因表达的影响至关重要.
研究的目的:
- 开发一种多重复合,高通量记者分析,用于对转录活动的并行监测.
- 调查局部染色体环境和调控元素对基因表达的影响.
- 分析大型数据集的记者集成,以获得对基因调节的新见解.
主要方法:
- 开发一种多重化方法,用于并行监控数千个报告员集成.
- 整合了超过27,000个不同的记者构造到小鼠胚胎干细胞中.
- 分析与基因组位置和染色体特征相关的记者表达水平.
主要成果:
- 在成千上万的记者集成中,表达级别的变化达到了约1000倍.
- 确定了与膜相关的域作为转录减弱剂.
- 显示的染色质紧缩是预测记者活性和估计增强剂影响的20 kb.
结论:
- 复杂记者测定使得基因调节的高通量分析成为可能.
- 染色体结构显著影响基因表达水平.
- 该试验为各种基因调节研究提供了一个灵活的平台.
相关概念视频
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...

