IMAGEtags: Quantifying mRNA Transcription in Real Time with Multiaptamer Reporters.
IMAGEtags: Quantifying mRNA Transcription in Real Time with Multiaptamer Reporters.
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DOI：
10.1016/bs.mie.2016.02.028
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发表时间：
2016
影响因子：
--
通讯作者：
Nilsen-Hamilton M
中科院分区：
文献类型：
--
作者：
Ray J;Shin I;Ilgu M;Bendickson L;Gupta V;Kraus GA;Nilsen-Hamilton M
细胞通讯对多细胞生物的组织、发育和维持至关重要。这种交流大部分涉及RNA转录的变化，并且是动态的。大多数研究转录的方法需要通过牺牲交流细胞来中断细胞功能的连续性，并通过定期采样单个细胞或群体来获取基因表达信息。IMAGEtag技术量化单个活细胞中的RNA水平，在酵母中展示，允许活细胞随着时间的推移被跟踪，因为它们对不同的环境线索作出反应。IMAGEtags是一种短rna，由一系列可变数量的串联适体组成，可以结合小分子配体。适配体串可以在长度和适配体成分的配置上变化，例如包含多个相同的适配体或两个或多个不同的适配体，这些适配体交替出现。最小有效长度约为5个适体。最大长度未定义。小分子配体可以成像为荧光基团缀合物。对于每个IMAGEtag，提供了两个荧光团共轭物，这是FRET对。当细胞表达含有IMAGEtag序列的RNA时，适体结合它们的配体，将荧光团带到足够近的距离以允许FRET。两个荧光团的背景荧光在FRET通道中是最小的。这些特点赋予了IMAGEtags报告活细胞mRNA表达水平的敏感性。
Cell communications are essential to the organization, development and maintenance of multicellular organisms. Much of this communication involves changes in RNA transcription and is dynamic. Most methods for studying transcription require interrupting the continuity of cellular function by sacrificing the communicating cells and capturing gene expression information by periodic sampling of individual cells or the population. The IMAGEtag technology to quantify RNA levels in individual living cells, demonstrated here in yeast, allows living cells to be tracked over time as they respond to different environmental cues. IMAGEtags are short RNAs consisting of strings of a variable number of tandem aptamers that bind small molecule ligands. The aptamer strings can vary in length and in configuration of aptamer constituents, such as to contain multiples of the same aptamer or two or more different aptamers that alternate in their occurrence. A minimum effective length is about 5 aptamers. The maximum length is undefined. The small molecule ligands are enabled for imaging as fluorophore conjugates. For each IMAGEtag, two fluorophore conjugates are provided, which are FRET pairs. When a cell expresses an RNA containing an IMAGEtag sequence, the aptamers bind their ligands and bring the fluorophores in to sufficiently close proximity to allow FRET. The background fluorescence of both fluorophores is minimal in the FRET channel. These features endow IMAGEtags with the sensitivity to report on mRNA expression levels in living cells.
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DOI：
10.1126/science.1207339
发表时间：
2011-07-29
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Paige JS;Wu KY;Jaffrey SR
通讯作者：
Jaffrey SR
影响因子：
4.6
作者：
Kraus, George A.;Jeon, Insik;Parvin, Bahram
通讯作者：
Parvin, Bahram
影响因子：
4.5
作者：
Hogg, J. Robert;Collins, Kathleen
通讯作者：
Collins, Kathleen
影响因子：
48
作者：
Strack, Rita L.;Disney, Matthew D.;Jaffrey, Samie R.
通讯作者：
Jaffrey, Samie R.
影响因子：
48
作者：
Hocine, Sami;Raymond, Pascal;Zenklusen, Daniel;Chao, Jeffrey A.;Singer, Robert H.
通讯作者：
Singer, Robert H.