HITS-CLIP为大脑的替代RNA处理提供了全基因组的洞察力
Donny D Licatalosi1, Aldo Mele, John J Fak
1Laboratory of Molecular Neuro-Oncology and Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.
Nature
|November 4, 2008
概括
通过交联免疫沉降 (HITS-CLIP) 隔离的RNA的高通量测序在体内映射蛋白质-RNA相互作用. 这种方法揭示了Nova拼接因子如何调节基因表达,包括大脑中的替代拼接和多化.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 神经科学是一个神经科学.
背景情况:
- 蛋白质-RNA相互作用对于基因表达至关重要.
- 使用生化方法在活体组织中研究这些相互作用是具有挑战性的.
研究的目的:
- 在体内开发一种全基因组的方法来绘制蛋白质-RNA结合点的地图.
- 研究神经元特异性拼接因子Nova的体内RNA结合场景.
主要方法:
- 通过交联免疫沉降 (HITS-CLIP) 隔离的RNA的高通量测序.
- 在小鼠大脑中对Nova-RNA相互作用的分析.
主要成果:
- HITS-CLIP在体内为Nova生成了可复制的全基因组RNA结合图.
- 这些地图证实了Nova在确定替代拼接结果中的作用,并预测了其 de novo 的作用.
- 在3'未翻译区域中确定了许多Nova-RNA相互作用,揭示了Nova在大脑中替代多基解的调节.
结论:
- HITS-CLIP是一种强大而公正的方法,用于在体内识别功能性蛋白质-RNA相互作用.
- 这种技术为通过像Nova.com这样的拼接因子调节基因表达提供了新的见解.
相关概念视频
Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
RNA-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...
Ribosome 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...
Pre-mRNA Processing: RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...


