Top > Search of International Patents > CONSTRUCTION OF PROTEIN-RESPONSIVE shRNA/RNAi REGULATION SYSTEM UTILIZING RNP MOTIF

CONSTRUCTION OF PROTEIN-RESPONSIVE shRNA/RNAi REGULATION SYSTEM UTILIZING RNP MOTIF

Foreign code F120006324
File No. I029P008WO
Posted date Mar 22, 2012
Country EPO
Application number 09831915
Gazette No. 2390324
Gazette No. 2390324
Date of filing Dec 9, 2009
Gazette Date Nov 30, 2011
Gazette Date Oct 1, 2014
International application number JP2009070580
International publication number WO2010067811
Date of international filing Dec 9, 2009
Date of international publication Jun 17, 2010
Priority data
  • 2009JP070580 (Dec 9, 2009) WO
  • P2008-312951 (Dec 9, 2008) JP
Title CONSTRUCTION OF PROTEIN-RESPONSIVE shRNA/RNAi REGULATION SYSTEM UTILIZING RNP MOTIF
Abstract An object of the present invention is to provide an RNAi control system using an RNA-protein interaction motif.
The present invention provides an shRNA comprising: a guide strand having a sequence complementary to a target sequence; a passenger strand which forms a duplex with the guide strand; and a linker strand which links the guide strand and the passenger strand, wherein the linker strand comprises an RNP-derived protein-binding motif sequence.
The present invention also provides an RNAi control system comprising: the shRNA; and an RNP-derived protein which specifically binds to a protein-binding motif sequence in the shRNA.
Scope of claims [claim1]
1. A RNAi control system responsive to a protein expressed in a cell, the system comprising: a vector for expression of a shRNA comprising: a guide strand having a sequence complementary to an mRNA of a target sequence; a passenger strand which forms a duplex with the guide strand; and a linker strand which links the guide strand and the passenger strand, the linker strand comprising a Box CD sequence, wherein the binding of a L7Ae protein or L7Ae protein-containing fusion protein expressed in the cell to the shRNA inhibits the cleavage of the shRNA by Dicer.
[claim2]
2. An in vitro RNAi control method responsive to a protein expressed in a cell, the method comprising the step of: introducing into the cell a vector for expression of a shRNA comprising a guide strand having a sequence complementary to an mRNA of a target sequence; a passenger strand which forms a duplex with the guide strand; and a linker strand which links the guide strand and the passenger strand, the linker strand comprising a Box CD sequence, wherein the binding of a L7Ae protein or L7Ae protein-containing fusion protein expressed in the cell to the shRNA inhibits the cleavage of the shRNA by Dicer.
[claim3]
3. The RNAi control system according to claim 1, wherein the target sequence of the shRNA is Bcl-xL mRNA, and the RNAi control system controls the expression of an apoptosis regulatory protein.
[claim4]
4. A shRNA comprising: a guide strand having a sequence complementary to a mRNA of a target sequence; a passenger strand which forms a duplex with the guide strand; and a linker strand which links the guide strand and the passenger strand, the linker strand comprising a Box CD sequence,
wherein in response to a L7Ae protein or L7Ae protein-containing fusion protein expressed in a cell, the cleavage of the shRNA by Dicer is inhibited to control the expression of a protein encoded by the mRNA of the target sequence.
[claim5]
5. The RNAi control system according to claim 1, further comprising a vector for intracellular expression of the L7Ae protein or L7Ae protein-containing fusion protein.
[claim6]
6. The RNAi control system according to claim 1, wherein the target sequence of the shRNA is GFP mRNA.
[claim7]
7. The in vitro RNAi control method according to claim 2, wherein the target sequence of the shRNA is Bcl-xL mRNA, and the RNAi control method controls the expression of an apoptosis regulatory protein.
[claim8]
8. The in vitro RNAi control method according to claim 2, further comprising the step of introducing a vector for intracellular expression of the L7Ae protein or L7Ae protein-containing fusion protein into the cell.
[claim9]
9. A method for quantifying the expression of an intracellular marker protein comprising an L7Ae protein without destroying a cell, comprising the steps of: introducing a shRNA comprising: a guide strand having a sequence complementary to GFP mRNA; a passenger strand which forms a duplex with the guide strand; and a linker strand which links the guide strand and the passenger strand, the linker strand comprising a Box CD sequence; and measuring the fluorescence intensity of the GFP.
  • Applicant
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • Inventor
  • INOUE TAN
  • SAITO HIROHIDE
  • KASHIDA SHUNICHI
  • HAYASHI KARIN
IPC(International Patent Classification)
U.S. Cl./(Sub)
  • C12N015/11M
  • M12N310/14
  • M12N310/3519
  • M12N310/531
  • M12N320/50
Specified countries Contracting States: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO SE SI SK SM TR
Please contact us by E-mail or facsimile if you have any interests on this patent.

PAGE TOP

close
close
close
close
close
close