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FLUOROGENIC NUCLEIC ACID MOLECULE AND TARGET-RNA FLUORESCENT LABELING METHOD

外国特許コード F200010216
整理番号 AF46-02WO
掲載日 2020年8月4日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2019JP047228
国際公開番号 WO 2020116446
国際出願日 令和元年12月3日(2019.12.3)
国際公開日 令和2年6月11日(2020.6.11)
優先権データ
  • 特願2018-226743 (2018.12.3) JP
発明の名称 (英語) FLUOROGENIC NUCLEIC ACID MOLECULE AND TARGET-RNA FLUORESCENT LABELING METHOD
発明の概要(英語) The present invention provides: a fluorogenic RNA capable of visualizing mRNA in the cell, in particular, in a living cell of a mammal; and a target-RNA fluorescent labeling method utilizing the same. The present invention provides a fluorogenic nucleic acid molecule characterized in that: a base sequence in which two or more fluorescent-molecule binding regions are linked via a linker sequence is contained; and the fluorescent-molecule binding regions each have one or more fluorescent-molecule binding aptamer sequences inserted into a scaffold sequence.
従来技術、競合技術の概要(英語) BACKGROUND ART
Gene expression level of mRNA which is controlled for a better understanding, a space-time kinetics of the mRNA in living cells the establishment of a method to visualize is strongly demanded. Visualization of mRNA in living cells of a general method is used, the target mRNA, such as MS2 or PP713 RNA stem, loop tag is added, and the fluorescent molecule is a method to recruit the fusion protein. The art, in the cytoplasm or track the motion of the mRNA (for example, see Non-Patent Document 1.), The proteins in the nucleus in order to visualize mRNA (for example, see Non-Patent Document 2.), Are applied. However, for the tag in addition to mRNA, mRNA in a cell or to visualize the dynamics of the distribution, when the level of expression, often, a high background of unbound fluorescence protein, interpretation of the results difficult.
MRNA in living cells as a method of visualization, a particular target molecule binds to the nucleotide sequence-dependent RNA aptamer is mentioned as a candidate method. RNA aptamers are of certain, low-molecular-weight compound that binds the fluorescent and the fluorescence can be enhanced by (for example, see Non-Patent Document 3 and 4.). RNA aptamers such as, fluorogenic RNA(fluorogenic RNA) are also referred. Fluorogenic RNA may include, for example fluorophores, GFP(Green fluorescent protein) DFHBI that are analogs of the family of low molecular weight (3,5-hydroxy benzylidene imidazolinone difluoro-4-) and the fluorescent molecule of the binding and Broccoli Spinach, and derivatives thereof (for example, Non-Patent Document 5, Patent Document 1 the reference.).
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • OKADA Yasushi
  • ARIYOSHI Tetsuro
国際特許分類(IPC)
参考情報 (研究プロジェクト等) CREST Innovative Technology Platforms for Integrated Single Cell Analysis AREA
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