在Drosophila RNA干扰查中,非目标效应的发生率
Yong Ma1, Adrian Creanga, Lawrence Lum
1Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Nature
|September 12, 2006
概括
在RNA干扰 (RNAi) 选中,非目标效应 (OTEs) 在Drosophila中很常见,即使是长长的双链RNA (dsRNAs). 基因和dSRNA中的并列重复经常会在全基因组RNAi屏幕中导致错误阳性.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物化学 生物化学
背景情况:
- RNA干扰 (RNAi) 使用小RNA来调节基因表达.
- 目标外效应 (OTEs) 可能来自小RNA和意想不到的目标之间的不完美合.
- 长长的双链RNAs (dsRNAs) 被认为通过细胞内加工成小RNAs来减少OTEs.
研究的目的:
- 在Drosophila的全基因组RNAi查中调查OTEs的流行率.
- 为了识别无翼 (Wg) 信号传导的新组件.
- 为了解决RNAi选方法的可靠性.
主要方法:
- 在Drosophila S2R+细胞中全基因组RNAi屏幕.
- 无翼 (Wg) 信号传输路径组件的分析.
- 识别和描述非目标效应.
主要成果:
- 在Drosophila中,长dRNAs表现出由短的同源性延伸介导的普遍的OTEs.
- 在选中,很少有Wg途径组件的合法候选者被确定.
- 发现dSRNA和基因中的协同三核酸重复会导致乱交的OTE和错误阳性.
结论:
- 在Drosophila RNAi查中,OTE是一个重要的问题,影响了真正的生物标的识别.
- 串联重复是全基因组RNAi查中错误阳性的常见来源.
- 实施减轻OTE的措施对于提高Drosophila和其他生物体RNAi查可靠性至关重要.
相关概念视频
01:23RNA 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...
02:46Genetic Screens
Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...
02:15Experimental 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...