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1. WO2021010442 - PROTEIN AND RNA INTERACTION EVALUATION METHOD, INTERACTION MODULATOR EVALUATION METHOD AND INTERACTION MODULATOR DETECTION METHOD, AND FUSION PROTEIN, KIT AND BIOSENSOR USING SAME

Publication Number WO/2021/010442
Publication Date 21.01.2021
International Application No. PCT/JP2020/027710
International Filing Date 16.07.2020
IPC
C12N 15/11 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
C12Q 1/68 2018.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
G01N 33/50 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
G01N 33/68 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
68involving proteins, peptides or amino acids
CPC
C12N 15/11
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
C12Q 1/68
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
G01N 33/50
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
G01N 33/68
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
68involving proteins, peptides or amino acids
Applicants
  • 国立大学法人東京大学 THE UNIVERSITY OF TOKYO [JP]/[JP]
Inventors
  • 遠藤 慧 ENDO Kei
  • 伊藤 耕一 ITO Koichi
Agents
  • 特許業務法人セントクレスト国際特許事務所 CENTCREST IP ATTORNEYS
Priority Data
2019-13119816.07.2019JP
Publication Language Japanese (JA)
Filing Language Japanese (JA)
Designated States
Title
(EN) PROTEIN AND RNA INTERACTION EVALUATION METHOD, INTERACTION MODULATOR EVALUATION METHOD AND INTERACTION MODULATOR DETECTION METHOD, AND FUSION PROTEIN, KIT AND BIOSENSOR USING SAME
(FR) PROCÉDÉ D'ÉVALUATION D'INTERACTION ENTRE PROTÉINE ET ARN, PROCÉDÉ D'ÉVALUATION DE MODULATEUR D'INTERACTION ET PROCÉDÉ DE DÉTECTION DE MODULATEUR D'INTERACTION, ET PROTÉINE DE FUSION, KIT ET BIOCAPTEUR L'UTILISANT
(JA) タンパク質とRNAとの相互作用評価方法、相互作用調節物質評価方法、及び相互作用調節物質検出方法、並びに、これらに用いる融合タンパク質、キット、及びバイオセンサ
Abstract
(EN)
Provided is a method for evaluating the interaction of a protein and RNA, said method comprising: (1) a step for bringing the following (a), (b), and (c) into contact in cytoplasm or a cell-free system, in which (a) is a first fusion protein obtained by fusing a subject protein and a first element of a marker protein, (b) is a second fusion protein obtained by fusing an RNA binding protein and a second element of the marker protein, and (c) is a ribonucleoprotein comprising fusion RNA which is obtained by fusing a subject RNA and RNA which forms a composite with the RNA binding protein; (2) a step for detecting a signal caused by the marker protein, which expresses a function by means of the first element and the second element being in close proximity; and (3) a step for determining the interaction of the subject protein and the subject RNA by means of detecting the signal, wherein if the subject protein and the subject RNA interacted, the first fusion protein and the ribonucleoprotein form a composite, and the marker protein function is expressed by means of the first element and the second element being in close proximity.
(FR)
L'invention concerne un procédé d'évaluation de l'interaction entre une protéine et un ARN, ledit procédé comprenant : (1) la mise en contact des éléments suivants (a), (b), et (c) dans le cytoplasme ou dans un système acellulaire, dans lequel (a) est une première protéine de fusion obtenue par fusion d'une protéine sujet et d'un premier élément d'une protéine marqueur, (b) est une seconde protéine de fusion obtenue par fusion d'une protéine de liaison à l'ARN et d'un second élément de la protéine marqueur, et (c) est une ribonucléoprotéine comprenant un ARN de fusion étant obtenu par fusion d'un ARN et d'ARN sujet formant un composite avec la protéine de liaison à l'ARN; (2) la détection d'un signal provoqué par la protéine marqueur, exprimant une fonction au moyen du premier élément et du second élément étant à proximité étroite; et (3) la détermination de l'interaction entre la protéine sujet et l'ARN sujet au moyen de la détection du signal, si la protéine de sujet et l'ARN sujet ont interagi, la première protéine de fusion et la ribonucléoprotéine forment un composite, et la fonction de protéine marqueur est exprimée au moyen du premier élément et du second élément étant à proximité étroite.
(JA)
タンパク質とRNAとの相互作用を評価する方法であって、 (1)細胞質又は無細胞系において、 (a)被検タンパク質とマーカータンパク質の第1の要素とが融合されてなる第1の融合タンパク質と、 (b)RNA結合性タンパク質と前記マーカータンパク質の第2の要素とが融合されてなる第2の融合タンパク質、及び(c)前記RNA結合性タンパク質と複合体を形成するRNAと被検RNAとが融合されてなる融合RNA、からなるリボヌクレオタンパク質と、 を接触させる工程、 (2)前記第1の要素と前記第2の要素との近接によって機能を発現した前記マーカータンパク質に起因するシグナルを検出する工程、並びに、 (3)前記シグナルの検出により、前記被検タンパク質と前記被検RNAとの相互作用を判定する工程、 を含み、 前記被検タンパク質と前記被検RNAとが相互作用した場合に、前記第1の融合タンパク質と前記リボヌクレオタンパク質とが複合体を形成し、かつ、前記第1の要素と前記第2の要素とが近接することにより前記マーカータンパク質の機能が発現する、方法。
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