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Protein-responsive translational regulatory system using RNA-protein interacting motif

Foreign code F110003837
File No. I029P004WO
Posted date Jul 6, 2011
Country United States of America
Application number 74385308
Gazette No. 20110040077
Gazette No. 9255274
Date of filing Nov 21, 2008
Gazette Date Feb 17, 2011
Gazette Date Feb 9, 2016
International application number JP2008071213
International publication number WO2009066757
Date of international filing Nov 21, 2008
Date of international publication May 28, 2009
Priority data
  • P2007-303662 (Nov 22, 2007) JP
  • P2008-186385 (Jul 17, 2008) JP
  • 2008WO-JP71213 (Nov 21, 2008) WO
Title Protein-responsive translational regulatory system using RNA-protein interacting motif
Abstract (US9255274)
An object of the present invention is to provide a translationally regulatable mRNA which has wider application and can perform specific ON-OFF regulation, an RNA-protein complex specifically bound to the mRNA, and a translational regulatory system.
The present invention provides an mRNA having an RNA-protein complex interacting motif-derived nucleotide sequence 5′ to the ribosome-binding site or within the 5′ region of the open reading frame, and an mRNA having a nucleotide sequence complementary to an RNA-protein complex interacting motif-derived nucleotide sequence 5′ to the ribosome-binding site or within the 5′ region of the open reading frame.
Scope of claims [claim1]
1. An isolated non-naturally occurring mRNA encoding a protein comprising an RNA-protein complex interacting motif nucleotide sequence incorporated 5' to a ribosome-binding site in a position 2 to 10 bases distant from the ribosome-binding site or within the 5' region of an open reading frame, wherein the interacting motif comprises nucleic acid sequence SEQ ID NO:9, wherein an RNA-protein complex of the interacting motif and L7Ae protein has a dissociation constant Kd of approximately 0.1 nM to approximately 1 mu M between the motif and L7Ae and the interacting motif interacts with L7Ae protein.
[claim2]
2. An RNA-protein complex comprising an mRNA according to claim 1 and a protein specifically binding to the nucleotide sequence.
[claim3]
3. A translational regulatory kit comprising an mRNA according to claim 1 and a protein specifically binding to the nucleotide sequence.
[claim4]
4. A method for translational regulation of mRNA, comprising contacting the mRNA according to claim 1 with a protein specifically binding to the RNA-protein complex interacting motif nucleotide sequence.
[claim5]
5. An artificial information conversion method which converts input information of an arbitrary substrate protein to output information of an arbitrary target protein using an mRNA according to claim 1, comprising steps of preparing the mRNA of claim 1 having an open reading frame encoding the arbitrary target protein; and
contacting the mRNA with the substrate protein that specifically binds to the RNA-protein complex interacting motif nucleotide sequence.
[claim6]
6. A plasmid vector comprising a nucleic acid sequence encoding an mRNA according to claim 1.
[claim7]
7. An intracellular translational regulatory kit comprising a first plasmid vector comprising a nucleic acid sequence encoding an mRNA according to claim 1, and
a second plasmid vector comprising a nucleic acid sequence encoding a protein specifically binding to the RNA-protein complex interacting motif nucleotide sequence.
[claim8]
8. The kit according to claim 7, for regulating protein translation in a human cancer cell.
  • Inventor, and Inventor/Applicant
  • INOUE TAN
  • SAITO HIROHIDE
  • KOBAYASHI TETSUHIRO
  • HARA TOMOAKI
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
U.S. Cl./(Sub)
  • C12N015/67
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