EP 1812582 A4 20130320 - SINGLE PROTEIN PRODUCTION IN LIVING CELLS FACILITATED BY A MESSENGER RNA INTERFERASE
Title (en)
SINGLE PROTEIN PRODUCTION IN LIVING CELLS FACILITATED BY A MESSENGER RNA INTERFERASE
Title (de)
EINZELPROTEINHERSTELLUNG IN LEBENDEN ZELLEN MITTELS MESSENGER-RNA-INTERFERASE
Title (fr)
PRODUCTION D'UNE SEULE PROTEINE DANS DES CELLULES VIVANTES FACILITEE PAR UNE INTERFERASE D'ARN MESSAGER
Publication
Application
Priority
- US 2005040107 W 20051104
- US 62497604 P 20041104
Abstract (en)
[origin: WO2006055292A2] 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.
IPC 8 full level
C12P 21/02 (2006.01); C12N 9/22 (2006.01)
CPC (source: EP KR)
C12N 5/00 (2013.01 - KR); C12N 5/10 (2013.01 - KR); C12N 9/22 (2013.01 - EP); C12P 21/02 (2013.01 - EP); C12P 21/06 (2013.01 - KR)
Citation (search report)
- [AP] WO 2004113498 A2 20041229 - UNIV NEW JERSEY MED [US], et al
- [XP] SUZUKI MOTOO ET AL: "Single protein production in living cells facilitated by an mRNA interferase", MOLECULAR CELL, CELL PRESS, CAMBRIDGE, MA, US, vol. 18, no. 2, 15 April 2005 (2005-04-15), pages 253 - 261, XP002545106, ISSN: 1097-2765, DOI: 10.1016/J.MOLCEL.2005.03.011
- [I] CHRISTENSEN S K ET AL: "Toxin-antitoxin Loci as Stress-response-elements: ChpAK/MazF and ChpBK Cleave Translated RNAs and are Counteracted by tmRNA", JOURNAL OF MOLECULAR BIOLOGY, ACADEMIC PRESS, UNITED KINGDOM, vol. 332, no. 4, 26 September 2003 (2003-09-26), pages 809 - 819, XP004454294, ISSN: 0022-2836, DOI: 10.1016/S0022-2836(03)00922-7
- See references of WO 2006055292A2
Designated contracting state (EPC)
AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI SK TR
DOCDB simple family (publication)
WO 2006055292 A2 20060526; WO 2006055292 A3 20060803; CA 2577180 A1 20060526; CN 101052713 A 20071010; CN 101052713 B 20110126; EP 1812582 A2 20070801; EP 1812582 A4 20130320; JP 2008518623 A 20080605; JP 5013375 B2 20120829; KR 101064783 B1 20110914; KR 20080088350 A 20081002
DOCDB simple family (application)
US 2005040107 W 20051104; CA 2577180 A 20051104; CN 200580032378 A 20051104; EP 05851377 A 20051104; JP 2007540092 A 20051104; KR 20077012576 A 20051104