Single-molecule analysis of gene expression using two-color RNA labeling in live yeast.

Single-molecule analysis of gene expression using two-color RNA labeling in live yeast.
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DOI:
10.1038/nmeth.2305
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发表时间:
2013-02
期刊:
影响因子:
48
通讯作者:
Singer, Robert H.
Singer, Robert H.
中科院分区:
生物学1区
文献类型:
--
作者:
Hocine, Sami;Raymond, Pascal;Zenklusen, Daniel;Chao, Jeffrey A.;Singer, Robert H.

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mRNA的活细胞成像为基因表达提供了重要的见解,但它通常仅限于一种RNA的标记,从未用于酵母中单个mRNA的计数。我们展示了一种使用MS2和PP7 RNA标记的单分子分辨率双色成像系统。我们使用这种方法来测量mRNA水平的内在噪声和单个基因的RNA聚合酶II动力学。
Live-cell imaging of mRNA yields important insights into gene expression, but it has generally been limited to the labeling of one RNA species and has never been used to count single mRNAs over time in yeast. We demonstrate a two-color imaging system with single-molecule resolution using MS2 and PP7 RNA labeling. We use this methodology to measure intrinsic noise in mRNA levels and RNA polymerase II kinetics at a single gene.
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