通过转基因RNAi对Drosophila中Notch信号的全基因组分析
Jennifer L Mummery-Widmer1, Masakazu Yamazaki, Thomas Stoeger
1Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr Bohr-Gasse 3.
Nature
|April 14, 2009
概括
这项研究使用了Drosophila的全基因组RNA干扰 (RNAi) 屏幕来分析感官器官发育中的Notch信号通路. 研究人员发现了调节这种途径和细胞分裂的新基因,为基因组研究创造了宝贵的资源.
科学领域:
- 发展生物学 发展生物学
- 基因组学就是基因组学.
- 细胞信号传递 细胞信号传递
背景情况:
- 全基因组RNA干扰 (RNAi) 屏幕是识别参与细胞过程的基因的强大工具.
- 之前的RNAi屏幕应用并没有广泛探索复杂的,组织特异性的发育过程.
- 诺奇信号通路在细胞命运的决定和发育中起着至关重要的作用.
研究的目的:
- 使用全基因组RNAi屏幕研究Drosophila在外部感官器官发育期间的Notch信号通路.
- 识别调节Notch信号和不对称细胞分裂的新型基因.
- 为Notch信号和不对称的细胞分裂构建一个功能互动网络.
主要方法:
- 利用了Drosophila菌株的库,使用可诱导的头RNAi结构来对向基因进行淘汰.
- 进行全基因组查,将假定的功能丧失表型分配给蛋白质编码基因.
- 采用了二次测试来验证基因功能,并将表型数据与蛋白质相互作用数据集成在一起.
- 应用集群算法来识别监管途径和复杂.
主要成果:
- 将假定功能丧失的表型分配给Drosophila蛋白质编码基因的21.2%.
- 发现了6个参与非对称细胞分裂的新基因.
- 发现了23种调节Notch信号通路的新基因.
- 为Notch信号和不对称细胞分裂构建了一个全基因组功能验证的相互作用网络.
- 确定了核进口通路和COP9信号作为Notch信号的调节器.
结论:
- 通过使用RNAi屏幕在全基因组范围内分析复杂的发育过程的可行性.
- 为Drosophila基因组的功能注释提供了一个全面的资源.
- 在发育背景下阐明了调节Notch信号和不对称细胞分裂的新组件和途径.
相关概念视频
03:14Notch Signaling Pathway
The Notch signaling pathway is a major intracellular signaling pathway that is highly conserved over a broad spectrum of metazoan species. It stands unique from other intracellular signaling mechanisms in animals because notch protein itself acts as the receptor as well as the primary signaling molecule.
The Notch gene came into the limelight in 1914 after the discovery that its mutation in Drosophila melanogaster leads to a serrated (or "notched") wing margin phenotype. It was not until 1985...
The Notch gene came into the limelight in 1914 after the discovery that its mutation in Drosophila melanogaster leads to a serrated (or "notched") wing margin phenotype. It was not until 1985...
03:14Notch Signaling Pathway
The Notch signaling pathway is a major intracellular signaling pathway that is highly conserved over a broad spectrum of metazoan species. It stands unique from other intracellular signaling mechanisms in animals because notch protein itself acts as the receptor as well as the primary signaling molecule.
The Notch gene came into the limelight in 1914 after the discovery that its mutation in Drosophila melanogaster leads to a serrated (or "notched") wing margin phenotype. It was not until 1985...
The Notch gene came into the limelight in 1914 after the discovery that its mutation in Drosophila melanogaster leads to a serrated (or "notched") wing margin phenotype. It was not until 1985...
01:12Role Of Notch Signalling In Intestinal Stem Cell Renewal
Notch signaling was first discovered in Drosophila melanogaster, where it is involved in cell lineage differentiation. Notch signaling regulates the maintenance and differentiation of intestinal stem cells or ISCs by controlling the expression of atonal homolog 1 or Atoh1. Atoh1 directs cells to differentiate into secretory cells.
Direct cell-to-cell contact is needed for the activation of Notch signaling. The signal is initiated when a notch ligand binds to a receptor on an adjacent cell, also...
Direct cell-to-cell contact is needed for the activation of Notch signaling. The signal is initiated when a notch ligand binds to a receptor on an adjacent cell, also...
02:54Canonical Wnt Signaling Pathway
The gene encoding the main signaling molecules of the Wnt signaling pathways (the Wnt proteins) was discovered almost four decades ago by Nüsslein-Volhard and Wieschaus. They identified and originally named the gene "wingless" (wg) after a phenotype discovered during their landmark genetic screen in Drosophila for body pattern defects. At around the same time, another researcher named Harold Varmus found that a murine tumor virus activates the mammalian wg homolog, Int-1, which results in tumor...