Methods for attenuating dengue virus infection

• US 8,796,235 B2
• Filed: 02/23/2004
• Issued: 08/05/2014
• Est. Priority Date: 02/21/2003
• Status: Expired due to Fees

• Current Assignee: University of South Florida (State University System of Florida)
• Original Assignee: University of South Florida (State University System of Florida)
• Inventors: Mohapatra, Shyam S., Zhang, Weidong
• Primary Examiner(s): SCHNIZER, RICHARD A
• Application Number:
  US10/544,146
• Publication Number:
  US 20060239971A1
• Time in Patent Office: 3,816 Days
• Field of Search: None
• US Class Current: 514/44.A
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61K 48/0058 Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/1131 against viruses
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed
• CPC Class Codes: C12N 2330/30 chemically synthesised
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention pertains to vectors for regulating gene expression having at least one gene expressing cassette and at least one gene suppressing cassette, wherein the gene expression cassette encodes a polypeptide of interest, and wherein the gene suppressing cassette encodes a short interfering RNA (siRNA) molecule that reduces expression of a target gene by RNA interference. The present invention further includes vectors that contain suppressor cassettes in conjunction with cassettes upregulating gene expression regulated by either a constitutive promoter, such as a general CMV promoter, or a tissue specific promoter. The present invention further includes vectors that contain Dengue virus gene suppression cassettes. The present invention further includes pharmaceutical compositions containing such vectors, methods of modulating the expression of genes in a host using such vectors, and method of producing such vectors.

Claims

1. A method for attenuating Dengue virus (DV) infection in human cells susceptible to DV infection in vivo, said method comprising administering to the cells in vivo an effective amount of a vector comprising a gene suppressing cassette, wherein the gene suppressing cassette comprises a polynucleotide operably-linked to a promoter sequence, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO:

4, wherein the polynucleotide encodes a short interfering RNA (siRNA) molecule that targets a sequence within the 3′

non-coding region of the DV genome that is common to four serotypes of DV, and wherein the polynucleotide is transcribed to produce the siRNA molecule.

2. The method of claim 1, wherein the vector is conjugated with chitosan.

3. The method of claim 1, wherein the vector is a non-viral vector.

4. The method of claim 1, wherein the vector is a viral vector.

5. The method of claim 1, wherein the vector is an adenoviral vector or adeno-associated viral vector.

6. The method of claim 1, wherein the vector is an adeno-associated viral vector.

7. The method of claim 1, wherein the siRNA molecule attenuates DV replication in the cells.

8. The method of claim 1, wherein the siRNA molecule has a hairpin structure.