在单核酸分辨率下调查的真核细胞转录组的动态谱
Brian T Wilhelm1, Samuel Marguerat, Stephen Watt
1Cancer Research UK Fission Yeast Functional Genomics Group, Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1HH, UK.
Nature
|May 20, 2008
概括
高通量测序揭示了裂变酵母中广泛的转录复杂性,揭示了新型RNA和各种条件下的动态拼接模式. 这种先进的RNA-Seq方法提供了比以前的技术更深入,更准确的转录组视图.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 文字转录学 (Transcriptomics) 是一个学科.
背景情况:
- 基因组研究表明,转录区域比以前理解的要复杂得多.
- 功能转录性质通常来自未翻译区域或非编码RNA中的调节序列,而不仅仅是编码序列.
- 以前的转录组研究依赖于杂交方法,限制了范围和条件.
研究的目的:
- 全球采样和分析裂变酵母Schizosaccharomyces pombe的转录组.
- 揭示各种环境,发育和遗传因素下的转录景观的动态可塑性.
- 通过识别新的转录和基因结构来改进现有的基因组注释.
主要方法:
- 采用了对互补DNA的直接高通量测序 (RNA-Seq).
- 补充了序列数据与高密度的瓦阵列.
- 在多种条件下的受访转录组:快速增殖,介质分化,环境压力和RNA处理突变.
主要成果:
- RNA-Seq提供了一种强大,定量和高分辨率的方法,用于以最小的背景噪音进行转录组采样.
- 检测到在基因组>90%的广泛转录,包括短暂或快速降解的RNA.
- 识别了许多新的非编码转录,未翻译的区域和基因结构,显著提高了基因组注释.
- 揭示了跨引子,基因和条件的拼接效率的惊人变化,与转录水平协调.
结论:
- 与杂交相比,高通量测序是深度和准确的转录组分析的优越方法.
- 裂变酵母转录组是高度动态和可塑的,受到环境,发育和遗传因素的影响.
- 调节的内子拼接广泛发生,与细胞增殖和分化有关.
相关概念视频
00:58Genomic DNA in Eukaryotes
Eukaryotes have large genomes compared to prokaryotes. To fit their genomes into a cell, eukaryotic DNA is packaged extraordinarily tightly inside the nucleus. To achieve this, DNA is tightly wound around proteins called histones, which are packaged into nucleosomes that are joined by linker DNA and coil into chromatin fibers. Additional fibrous proteins further compact the chromatin, which is recognizable as chromosomes during certain phases of cell division.
03:21RNA-seq
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
02:24Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...