阿尔戈纳特HITS-CLIP解码了微RNA-mRNA相互作用地图
Sung Wook Chi1, Julie B Zang, Aldo Mele
1Laboratory of Molecular Neuro-Oncology and Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.
Nature
|June 19, 2009
概括
通过交联免疫沉降 (HITS-CLIP) 隔离的RNA的高通量测序确定了小鼠大脑中的微RNA (miRNA) 和目标信使RNA (mRNA) 相互作用. 这种方法绘制了miRNA结合部位的地图,进步了对基因调节的理解.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 神经科学是一个神经科学.
背景情况:
- 微RNAs (miRNAs) 通过与信使RNAs (mRNAs) 的基因配对来调节基因表达.
- 预测miRNA-mRNA相互作用是具有挑战性的,因为结合所需的短序列 (6-8核酸) 是具有挑战性的.
- 通过交联免疫沉降 (HITS-CLIP) 隔离的RNA的高通量测序识别了功能性蛋白质-RNA相互作用.
研究的目的:
- 使用HITS-CLIP在体内绘制miRNA和mRNA结合部位的地图.
- 为了确定小鼠大脑中的特定miRNA-mRNA相互作用.
- 为探索miRNA作用和针对临床相关相互作用提供一个平台.
主要方法:
- 在小鼠大脑中的原生阿尔戈纳特 (Ago) 蛋白-RNA复合体的共价交联.
- 使用HITS-CLIP生成同时的Ago-miRNA和Ago-mRNA结合位点数据.
- 生物信息分析用于识别miRNA-mRNA相互作用部位.
- 对miR-124的相互作用图的全基因组验证和20种丰富的miRNA的映射.
主要成果:
- 从小鼠大脑生成的同时Ago-miRNA和Ago-mRNA结合部位数据.
- 确定了miR-124.4的全基因组相互作用图.
- 在P13小鼠大脑中创建了20个最丰富的miRNA的相互作用地图.
- 建立了miRNA-mRNA向的精确序列.
结论:
- 阿戈HITS-CLIP是一个多功能平台,用于研究miRNA的特异性和 in vivo 的功能.
- 这种方法可以识别精确的miRNA-mRNA相互作用.
- 这些发现有助于针对临床相关的miRNA-mRNA相互作用.
相关概念视频
MicroRNAs
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends...
MicroRNAs
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends...
MicroRNAs
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
RNA Interference
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
RNA Interference
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
Experimental RNAi
RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...


