Alnylam Pharmaceuticals, Inc.

American Home Products Corporation

Other

The General Hospital Corporation

Veritas Bio, LLC

Nucleonics, Inc.

Catherine Julia Pachuk

Chandrasekhar Satishchandran

Liat Mintz

Vincent R Zurawski

C Satishchandran

Daniel E McCallus

Richard B Ciccarelli

Leslie R Coney

Jack Raymond Wands

Takaji Wakita

Vincent R Zurawski, Jr

Tony Giordano

Manoj Samuel

Baohua Gu

Martin D Sigg, Jr

Fred T Oakes

Kathleen Herold

Daniel E Mccallus

Satishchandran C

Bachua Gu

Conserved consensus sequences from known hepatitis B virus strains and known hepatitis C virus strains, which are useful in inhibiting the expression of the viruses in mammalian cells, are provided. These sequences are useful to silence the genes of HBV and HCV, thereby providing therapeutic utility against HBV and HCV viral infection in humans.

Conserved HBV and HCV sequences useful for gene silencing

The invention relates to methods and compositions for modulating viral replication through double-stranded RNA-mediated gene silencing (RNAi), wherein the antiviral methods and compositions preferentially target opposite strand replication intermediates of single-stranded RNA viruses.

Targeting opposite strand replication intermediates of single-stranded viruses by RNAI

Described herein are methods for identifying nucleic acid sequences that modulate the function of a cell, the expression of a gene in a cell, or the biological activity of a target polypeptide in a cell. The methods involve the use of double stranded RNA expression libraries, double stranded RNA molecules, and post-transcriptional gene silencing techniques.

Post-transcriptional gene silencing using expressed double stranded RNA

The present invention provides agents, compositions, constructs and methods for silencing HCV polynucleotides, as well as methods and compositions for treating or preventing HCV infection in a mammalian cell. In one aspect, the present invention provides an agent or composition comprising at least one double-stranded RNA effector molecule or complex. The double-stranded RNA effector molecule or complex comprises: (1) a sequence of at least 19 nucleotides having at least 90% identity with a nucleotide sequence within HCV Conserved Region 1 (SEQ ID NO: 2), HCV Conserved Region 2 (SEQ ID NO: 3), HCV Conserved Region 5 (SEQ ID NO: 4), (ATR)-1 (SEQ ID NO: 86), ATR-2 (SEQ ID NO: 87), ATR-3 (SEQ ID NO: 88), ATR-4 (SEQ ID NO: 89); and (2) its complementary sequence. In another aspect, the present invention provides a construct suitable for replication in a host cell, and/or suitable for expression of an RNA molecule or complex of the invention in vitro or in vivo. In a third aspect, the present invention provides a method for silencing HCV RNA in a mammalian cell, which comprises administering to the mammalian cell an agent, composition, or construct of the invention in a manner and amount effective to silence HCV RNA in the cell. In a related aspect, the invention provides a method for treating or preventing HCV infection in a patient, comprising administering to the patient an effective amount of an agent, composition, or construct of the invention as described herein.

Hepatitis C dsRNA effector molecules, expression constructs, compositions, and methods of use

The invention encompasses methods of delivering nucleic acids, including dsRNA, to mammalian target cells in vivo via intercellular transfer, wherein the dsRNA is delivered to or expressed in a first cell different from the target cell, wherein the first cell facilitates delivery of the dsRNA to the target cell.

In vivo delivery of double stranded RNA to a target cell

The present invention provides methods of post-transcriptional gene silencing which involve the use of a first dsRNA having substantial sequence identity to a target nucleic acid and a short, second dsRNA which inhibits dsRNA-mediated toxicity. These methods can be used to prevent or treat a disease or infection by silencing a gene associated with the disease or infection. The invention also provides methods for identifying nucleic acid sequences that modulate a detectable phenotype, including the function of a cell, the expression of a gene, or the biological activity of a target polypeptide.

Methods and compositions for silencing genes without inducing toxicity

Expression constructs comprising at least two different RNA polymerase III promoters, wherein each promoter is operably linked to a nucleic acid sequence encoding an RNA effector molecule, are disclosed herein. Further provided are expression constructs comprising multiple polymerase III promoters operably linked to sequences encoding short hairpin RNA molecules, which may comprise single and/or multiple fingers. The provided constructs are useful for in vivo delivery of RNA molecules effective in gene silencing, including of viral genes including HBV and HCV.

Multiple RNA polymerase III promoter expression constructs

The invention features methods of analyzing the kinetics properties of transfection reactions. Also featured are methods for creating structural promoters which are effectively unregulated by enhancers and repressors. The structural promoters are significantly more active than the native promoter sequences upon which they are based.

Transfection kinetics and structural promoters

Nucleic acid molecule that comprise an incomplete hepatitis C viral genome are provided. Pharmaceutical compositions that contain nucleic acid molecules comprising an incomplete hepatitis C viral genome including a nucleotide sequence encoding a complete hepatitis C core protein operably linked to regulatory elements functional in human cells are provided. Methods of immunizing individuals susceptible to or infected by hepatitis C virus comprising the step of administering such pharmaceutical compositions are provided.

Growing community of inventors

North Wales, PA, United States of America

Catherine Julia Pachuk