Title:
SINGLE PROMOTER SYSTEM FOR MAKING SIRNA EXPRESSION CASSETTES AND EXPRESSION LIBRARIES USING A POLYMERASE PRIMER HAIRPIN LINKER
Document Type and Number:
WIPO Patent Application WO2004009796
Kind Code:
A3
Abstract:
The present invention relates to methods and compositions for the elucidation of gene function and the identification of novel genes. Specifically, the present invention relates to methods and compositions for improved functional genomic screening, functional inactivation of specific essential or non-essential genes, and identification of genes that are modulated in response to specific stimuli or encode recognizable phenotypic traits. In particular, the compositions of the present invention include, but are not limited to, expression cassettes comprising a novel polymerase primer hairpin linker that allows rapid construction of a single transcriptional unit encoding both strands of a hairpin siRNA, regardless of sequence. In addition, the present invention includes libraries comprising the expression cassettes of the invention, including vectors for transforming cells, such as replication-deficient retroviral vectors. The invention also includes methods for the production and screening of siRNA libraries, as well as therapeutic uses for the siRNAs expressed in accordance with the invention.
Inventors:
LI HENRY (US)
CHATTERTON JON E (US)
KE NING (US)
RHOADES KRISTINA L (US)
WONG-STAAL FLOSSIE (US)
CHATTERTON JON E (US)
KE NING (US)
RHOADES KRISTINA L (US)
WONG-STAAL FLOSSIE (US)
Application Number:
PCT/US2003/023239
Publication Date:
March 17, 2005
Filing Date:
July 23, 2003
Export Citation:
Assignee:
IMMUSOL INC (US)
LI HENRY (US)
CHATTERTON JON E (US)
KE NING (US)
RHOADES KRISTINA L (US)
WONG-STAAL FLOSSIE (US)
LI HENRY (US)
CHATTERTON JON E (US)
KE NING (US)
RHOADES KRISTINA L (US)
WONG-STAAL FLOSSIE (US)
International Classes:
C12N15/09; C12N15/11; C12N15/66; (IPC1-7): C12N15/63
Foreign References:
| US20040152172A1 | 2004-08-05 | |||
| US20040005593A1 | 2004-01-08 | |||
| US20030144239A1 | 2003-07-31 | |||
| US20040002077A1 | 2004-01-01 |
Other References:
MIYAGISHI ET AL.: "U6 promoter-driven siRNAs with four uridine 3'overhangs efficiently supress targeted gene expression in mammalian cells", NATURE BIOTECHNOLOGY, vol. 19, May 2002 (2002-05-01), pages 497 - 500, XP002965488
BRUMMELKAMP ET AL.: "A system for stable expression of short interfering RNAs in mammalian cells", SCIENCE, vol. 296, April 2002 (2002-04-01), pages 550 - 553, XP002234902
BEGER ET AL.: "Identification of Id4 as a regulator of BRCA1 expression by using a ribozyme library based inverse genomics approach", PROC. NATL. ACAD. SCI., vol. 98, January 2001 (2001-01-01), pages 130 - 135, XP002983351
KAWASAKI ET AL.: "Identification of genes by hybrid ribozymes that couple clevage activity with the unwinding activity of an endogenous RNA helicase", EMBO REPORTS, vol. 3, May 2002 (2002-05-01), pages 443 - 450, XP002983352
BRUMMELKAMP ET AL.: "A system for stable expression of short interfering RNAs in mammalian cells", SCIENCE, vol. 296, April 2002 (2002-04-01), pages 550 - 553, XP002234902
BEGER ET AL.: "Identification of Id4 as a regulator of BRCA1 expression by using a ribozyme library based inverse genomics approach", PROC. NATL. ACAD. SCI., vol. 98, January 2001 (2001-01-01), pages 130 - 135, XP002983351
KAWASAKI ET AL.: "Identification of genes by hybrid ribozymes that couple clevage activity with the unwinding activity of an endogenous RNA helicase", EMBO REPORTS, vol. 3, May 2002 (2002-05-01), pages 443 - 450, XP002983352
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