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DESIGN METHOD FOR RNA-BINDING PROTEIN USING PPR MOTIF, AND USE THEREOF 新技術説明会

外国特許コード F130007569
掲載日 2013年7月18日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2012JP077274
国際公開番号 WO 2013058404
国際出願日 平成24年10月22日(2012.10.22)
国際公開日 平成25年4月25日(2013.4.25)
優先権データ
  • 特願2011-231346 (2011.10.21) JP
発明の名称 (英語) DESIGN METHOD FOR RNA-BINDING PROTEIN USING PPR MOTIF, AND USE THEREOF 新技術説明会
発明の概要(英語) Provided is a method for designing a protein capable of selectively binding RNA bases or specifically binding an RNA base sequence. This protein includes 1 or more (preferably 2-14) PPR motifs comprising a polypeptide represented by formula (1) and having a length of 30-38 amino acids (in formula (1): Helix A is a moiety which is represented by formula (2), has a length of 12 amino acids, and is capable of forming an α-helix structure (in formula (2), A1-A12 each independently represent an amino acid); X is a moiety which is either not present or which has a length of 1-9 amino acids; Helix B is a moiety that has a length of 11-13 amino acids, and is capable of forming an α-helix structure; L is a moiety which is represented by formula (3) and has a length of 2-7 amino acids (in formula (3), each amino acid is numbered from the C-terminus in the following manner "i"(-1), "ii"(-2), on condition that in some cases Liii to Lvii are not present)). The protein corresponding to the RNA bases or base sequence that will be bound is obtained by combining: A1, A4, and 3 Lii amino acids; or A4 and 2 Lii amino acids.
従来技術、競合技術の概要(英語) BACKGROUND ART
In recent years, more various analysis revealed using nucleic acid binding protein factors, binds to the intended sequences established techniques, used. This sequence using the specific binding, the target nucleic acid (DNA or RNA) analysis of the subcellular localization of, the removal of the target DNA sequence, or downstream of the expression of the gene encoding the protein existing in the control (activation, or an inactivated) is becoming possible.
Factors acting on DNA as a protein, a zinc finger protein (non-patent document 1) or TAL effector (non-patent document 2, patent document 1) a protein engineering material research and development is performed, RNA specifically act on the development of protein factors are still very limited. This is because, in general the amino acid sequence constituting the protein having a binding affinity RNA RNA sequence and the law is almost not yet been clear, or not found to be due to the law. Exceptionally, puf made of amino acids 38 is repeatedly performed a plurality of motifs for pumilio protein, one of the puf 1 motif shown to bind to RNA 1 and the base (non-patent document 3), the binding characteristics of the new protein was used pumilio RNA protein with a, of the binding properties of modified RNA and techniques have been tried (non-patent document 4). However, puf and is highly conserved motif, and the existence of a small number of. Therefore, a limited RNA protein factors acting on the creation of the sequence is not used only.
On the other hand, from the genome sequence information, only as many as 500 plant protein to form a large family, a protein having protein (pentatricopeptide repeat (PPR) PPR motif) have been identified (non-patent document 5). PPR in nuclear-encoded proteins, organelles (chloroplasts and mitochondria) exclusively RNA level of control, cutting, translation, splicing, RNA editing, RNA stability which specifically act on the gene. PPR protein is, typically, 35 amino acid motif of low preservation property, i.e. about 10 contiguous nucleotides PPR motif has the structure, a combination of PPR motif, the sequence of the RNA to be responsible for the selective binding are considered to be. Most PPR PPR motif protein is about 10 repeat only and, in many cases, in order to exhibit the catalytic action domain necessary not found. Therefore, this PPR protein entities are believed adapter RNA (non-patent document 6).
The inventors have found that, using this PPR motif, for a method for modifying RNA-binding protein, has been proposed (patent document 2).
  • 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
  • NATIONAL UNIVERSITY CORPORATION KYUSHU UNIVERSITY
  • 発明者(英語)
  • NAKAMURA, Takahiro
  • YAGI. Yusuke
  • KOBAYASHI, Keiko
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 DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IS JP KE KG KM KN KP KR KZ LA LC LK LR LS LT 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 SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN 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 ML MR NE SN TD TG
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