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NUCLEIC ACID BINDING PROTEIN NEW

外国特許コード F210010309
整理番号 AF46-03WO
掲載日 2021年1月29日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2020JP015966
国際公開番号 WO 2020209332
国際出願日 令和2年4月9日(2020.4.9)
国際公開日 令和2年10月15日(2020.10.15)
優先権データ
  • 特願2019-074004 (2019.4.9) JP
発明の名称 (英語) NUCLEIC ACID BINDING PROTEIN NEW
発明の概要(英語) The present invention provides a novel modified protein which is to be used, as a novel detection tool relating to gene expression, for detecting a chromatin open structure more easily at a higher sensitivity than by the conventional technique. The present invention relates to: a nucleic acid binding fluorescent protein, said protein containing a DNA binding domain in which 3 or more TAL-repeats are repeatedly connected, characterized by binding independently from base sequences; and a method for fluorescent labeling of an open chromatin in a vital cell, said method comprising a step for transferring a gene encoding a nucleic acid binding protein into the vital cell, characterized in that the nucleic acid binding protein is a protein comprising a DNA binding domain, in which 3 or more TAL-repeats are repeatedly connected, and a fluorescent protein directly or indirectly bound thereto and the DNA binding domain binds to a nucleic acid independently from base sequences.
従来技術、競合技術の概要(英語) BACKGROUND ART
In recent years, attention has been paid to the results of the study on the mechanism of diversity of individual cells in spite of the same genome information. From the progress of this study, it was found that the maintenance of the essential genes and the suppression of the unnecessary genes are correctly regulated. The cell-specific trait appearing as a result of such control is maintained stably beyond cell division, and this event is called cell memory (or epigenetic).
TAL (Transcription Activator-Like) Effector (TALE) was identified as a transcription factor involved in gene expression regulation to suppress the immune mechanism of host cells when infected with pathogenic bacteria. TALE includes, at its center, a DNA binding domain consisting of a portion in which a structural unit (module) consisting of about 34 amino acids called TAL-repeat is linked by 10 ~ 30 repetitions. TAL-repeat consists of 2 set structures, where there are 2 amino acid residues involved in nucleobase recognition called Repeat Variable Diresidue (RVD) in the loop region connecting the 2 set structures. Individual TAL-repeat recognizes 1 nucleobases depending on the amino acid sequence of each RVD.
TALE can be modified so as to selectively bind to a nucleic acid containing a specific base sequence by linking TAL-repeat having RVD specifically recognizing a specific nucleic acid base in an appropriate order. That is, sequence-dependent binding can be made possible by using a module corresponding to the DNA sequence targeted for the DNA binding domain of TALE. For example, in genome editing, TALEN (TALE-Nuclease) in which TALE and FokI nuclease which recognize in dependence on the nucleic acid sequence are fused is used.
On the other hand, organisms have been known to have open chromatin structures by partially loosening aggregates of nucleic acids and histone in the nucleus in the regulation of expression of various genes in the development and differentiation. Analysis of whether or not the open chromatin structure takes place is particularly important in functional analysis of unidentified genes such as expression and identification of unknown differentiation-inducing genes involved in specific diseases, expression and identification of unidentified genes involved in carcinogenesis, and expression and identification of genes involved in metabolic abnormalities and neurodegeneration based on regulatory mechanisms which have not been elucidated yet.
  • 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
  • INO Daisuke
  • IKEDA Kazuho
国際特許分類(IPC)
指定国 National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JO JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN WS ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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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