WO2005023991A3 - Small hairpin rna libraries - Google Patents

Small hairpin rna libraries Download PDF

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Publication number
WO2005023991A3
WO2005023991A3 PCT/US2004/029176 US2004029176W WO2005023991A3 WO 2005023991 A3 WO2005023991 A3 WO 2005023991A3 US 2004029176 W US2004029176 W US 2004029176W WO 2005023991 A3 WO2005023991 A3 WO 2005023991A3
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WO
WIPO (PCT)
Prior art keywords
methods
hairpin rna
small hairpin
rna libraries
libraries
Prior art date
Application number
PCT/US2004/029176
Other languages
French (fr)
Other versions
WO2005023991A2 (en
Inventor
Alex Mccampbell
Aleksey G Kazantsev
Robert H Brown Jr
Anne B Young
Michele Maxwell
Original Assignee
Gen Hospital Corp
Alex Mccampbell
Aleksey G Kazantsev
Robert H Brown Jr
Anne B Young
Michele Maxwell
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Gen Hospital Corp, Alex Mccampbell, Aleksey G Kazantsev, Robert H Brown Jr, Anne B Young, Michele Maxwell filed Critical Gen Hospital Corp
Publication of WO2005023991A2 publication Critical patent/WO2005023991A2/en
Publication of WO2005023991A3 publication Critical patent/WO2005023991A3/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/111—General methods applicable to biologically active non-coding nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1093—General methods of preparing gene libraries, not provided for in other subgroups
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/11—Antisense
    • C12N2310/111—Antisense spanning the whole gene, or a large part of it
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering N.A.
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/50—Physical structure
    • C12N2310/53—Physical structure partially self-complementary or closed
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2320/00—Applications; Uses
    • C12N2320/10—Applications; Uses in screening processes
    • C12N2320/12—Applications; Uses in screening processes in functional genomics, i.e. for the determination of gene function
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2330/00—Production
    • C12N2330/30—Production chemically synthesised
    • C12N2330/31—Libraries, arrays

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The present invention relates to novel and rapid methods of identifying genes and pathways in mammalian cells based on RNA interference (RNAi). The invention includes methods of producing and using libraries of small interfering RNAs known as short hairpin RNAs (shRNAs). The dsRNAs generated by the methods of the invention can be introduced into cells in a number of ways, including as an isolated nucleic acid, (e.g., chemically synthesized or translated in vitro), or by transfection of an expression vector (e.g., a plasmid) that includes sequence encoding the shRNA.
PCT/US2004/029176 2003-09-05 2004-09-07 Small hairpin rna libraries WO2005023991A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US50086003P 2003-09-05 2003-09-05
US60/500,860 2003-09-05

Publications (2)

Publication Number Publication Date
WO2005023991A2 WO2005023991A2 (en) 2005-03-17
WO2005023991A3 true WO2005023991A3 (en) 2006-06-08

Family

ID=34273006

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2004/029176 WO2005023991A2 (en) 2003-09-05 2004-09-07 Small hairpin rna libraries

Country Status (1)

Country Link
WO (1) WO2005023991A2 (en)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2821085B1 (en) 2003-09-12 2020-04-29 University of Massachusetts Rna interference for the treatment of gain-of-function disorders
US8680063B2 (en) 2003-09-12 2014-03-25 University Of Massachusetts RNA interference for the treatment of gain-of-function disorders
US7498316B2 (en) 2004-04-06 2009-03-03 University Of Massachusetts Methods and compositions for treating gain-of-function disorders using RNA interference
JP2007274902A (en) * 2004-07-09 2007-10-25 Genofunction Inc METHOD FOR PRODUCING siRNA-EXPRESSING VECTOR LIBRARY
US20070161591A1 (en) 2005-08-18 2007-07-12 University Of Massachusetts Methods and compositions for treating neurological disease
US8735064B2 (en) 2007-12-24 2014-05-27 Bergenbio As Methods for creating and identifying functional RNA interference elements
GB0804690D0 (en) * 2008-03-13 2008-04-16 Netherlands Cancer Inst The Method
US7863222B2 (en) * 2008-08-01 2011-01-04 Rigel Pharmaceuticals, Inc. shRNA library
CA3216872A1 (en) * 2021-04-16 2022-10-20 Meso Scale Technologies, Llc. Methods, compositions, and kits for detecting hydrolase enzyme activity

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6586180B1 (en) * 1998-03-28 2003-07-01 University Of Utah Directed antisense libraries
US20030139363A1 (en) * 2001-07-23 2003-07-24 Kay Mark A. Methods and compositions for RNAi mediated inhibition of viral gene expression in mammals
US20030144239A1 (en) * 2001-12-24 2003-07-31 Reuven Agami Expression system

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6586180B1 (en) * 1998-03-28 2003-07-01 University Of Utah Directed antisense libraries
US20030139363A1 (en) * 2001-07-23 2003-07-24 Kay Mark A. Methods and compositions for RNAi mediated inhibition of viral gene expression in mammals
US20030144239A1 (en) * 2001-12-24 2003-07-31 Reuven Agami Expression system

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
DONZE TE AL: "RNA interference in mammalian cells using siRNAs synthesized with T7 RNA polymerase", NUCLEIC ACIDS RESEARCH, vol. 30, no. 10E46, 2002, pages 1 - 4, XP002973405 *
ELBASHIR ET AL: "RNA interference is mediated by 21-and 22-nucleotide RNAs", GENES AND DEVELOPMENT, vol. 15, 2002, pages 188 - 200, XP002206453 *
KAWASAKI ET AL: "siRNAs generated by recombinant human Dicer induce specific and significant but target site-independent gene silencing in human cells", NUCLEIC ACIDS RESEARCH, vol. 31, no. 3, 1 February 2003 (2003-02-01), pages 981 - 987, XP002971836 *
LAGOS-QUINTANA ET AL: "Identification of Novel Genes Coding for Small Expressed RNAs", SCIENCE, vol. 294, pages 853 - 858, XP002971837 *
MYERS ET AL: "Recombinant Dicer efficiently converts large dsRNAs into siRNAs suitable for gene silencing", NATURE BIOTECHNOLOGY, vol. 21, March 2003 (2003-03-01), pages 324 - 328, XP002302300 *
PFEFFER ET AL: "Cloning of small RNA molecules", CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, 2003, UNIT 26.4:26.4.1-26:4.18, XP008065837 *

Also Published As

Publication number Publication date
WO2005023991A2 (en) 2005-03-17

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