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Patent Searching and Data


Title:
SINGLE PROTEIN PRODUCTION IN LIVING CELLS FACILITATED BY A MESSENGER RNA INTERFERASE
Document Type and Number:
WIPO Patent Application WO2006055292
Kind Code:
A3
Abstract:
The present invention describes a single-protein production (SPP) system in living E. coli cells that exploits the unique properties of an mRNA interferase, for example, MazF, a bacterial toxin that is a single stranded RNA- and ACA-specific endoribonuclease, which efficiently and selectively degrades all cellular mRNAs in vivo, resulting in a precipitous drop in total protein synthesis. Concomitant expression of MazF and a target gene engineered to encode an ACA-less mRNA results in sustained and high-level (up to 90%) target expression in the virtual absence of background cellular protein synthesis. Remarkably, target synthesis continues for at least 4 days, indicating that cells retain transcriptional and translational competence despite their growth arrest. SPP technology works well for yeast and human proteins, even a bacterial integral membrane protein. This novel system enables unparalleled signal to noise ratios that should dramatically simplify structural and functional studies of previously intractable but biologically important proteins.

Inventors:
MASAYORI INOUYE (US)
ZHANG JUNJIE (CN)
SUZUKI MOTOO (US)
Application Number:
US2005040107W
Publication Date:
August 03, 2006
Filing Date:
November 04, 2005
Export Citation:
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Assignee:
UNIV NEW JERSEY MED (US)
MASAYORI INOUYE (US)
ZHANG JUNJIE (CN)
SUZUKI MOTOO (US)
International Classes:
C12N5/10; C12P21/00
Other References:
ZHANG Y. ET AL.: "MazF Cleaves Cellular mRNAs Specifically at ACA to Block Protein Synthesis in Escherichia coli", MOLECULAR CELL, vol. 12, October 2003 (2003-10-01), pages 913 - 923, XP002389064
ZHANG J. ET AL.: "Interference of mRNA Function by Sequence-specific Endoribonuclease PemK", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 279, no. 20, May 2004 (2004-05-01), pages 20678 - 20684, XP003001717
ZHANG Y. ET AL.: "Insights into the mRNA Cleavage Mechanism by MazF, an mRNA Interferase", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 280, no. 5, February 2005 (2005-02-01), pages 3143 - 3150, XP003001718
See also references of EP 1812582A4
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