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Small RNA-dependent translational regulatory system in cell or artificial cell model

Foreign code F170009178
File No. I029P002US1
Posted date Sep 12, 2017
Country United States of America
Application number 201314021260
Gazette No. 20140030707
Gazette No. 9057065
Date of filing Sep 9, 2013
Gazette Date Jan 30, 2014
Gazette Date Jun 16, 2015
International application number JP2008071214
International publication number WO2009066758
Date of international filing Nov 21, 2008
Date of international publication May 28, 2009
Priority data
  • P2007-303661 (Nov 22, 2007) JP
  • 2008WO-JP71214 (Nov 21, 2008) WO
  • 2010US-12743908 (Aug 31, 2010) US
Title Small RNA-dependent translational regulatory system in cell or artificial cell model
Abstract (US9057065)
An object of the present invention is to construct an mRNA which specifically responds to a short RNA sequence and can activate, repress, and regulate the translation of the desired gene, and to construct an artificial cell model system using a liposome comprising the mRNA and a cell-free translational system encapsulated therein.
The present invention provides: an mRNA comprising a target RNA-binding site located immediately 5′ to the ribosome-binding site, and a nucleotide sequence located 5′ to the target RNA-binding site, the nucleotide sequence being complementary to the ribosome-binding site; an mRNA comprising a small RNA-binding site located 3′ to the start codon, and a nucleotide sequence located 3′ to the small RNA-binding site, the nucleotide sequence encoding a protein; and a liposome comprising any of these mRNAs encapsulated therein.
Scope of claims [claim1]
1. An mRNA comprising a microRNA-binding site located 3' to the start codon and a nucleotide sequence located 3' to the microRNA-binding site, the nucleotide sequence encoding a protein, wherein the microRNA-binding site has a sequence complementary to a microRNA selected from the group consisting of miR164, miR170, miR171, miR156 miR159a and miR163.
[claim2]
2. A translation/expression regulation system comprising an mRNA according to claim 1.
[claim3]
3. A liposome comprising an mRNA according to claim 1 encapsulated therein.
[claim4]
4. The liposome according to claim 3, wherein the liposome is produced by the method comprising steps of: mixing one or more phospholipids, the mRNA, a cell-free translational system, and an aqueous solution into an oily liquid to form a W/O emulsion in which the mRNA and the cell-free translational system are encapsulated in the phospholipid vesicle;
adding an oily liquid containing outer membrane lipids dissolved therein, to an aqueous phase to form a molecular membrane in which the lipids are arranged at the oil/water interface; and
adding the W/O emulsion to the oil phase side of the interface and moving the W/O emulsion to the aqueous phase side of the interface such that the outer membrane lipid is added outside of the W/O emulsion to form a liposome.
  • Inventor, and Inventor/Applicant
  • INOUE TAN
  • SAITO HIROHIDE
  • KATO YUSHO
  • YOSHIKAWA KENICHI
  • YAMADA AYAKO
  • YAMANAKA TORU
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
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