Modulation of gene expression through interference with RNA secondary structure

• US 5,866,698 A
• Filed: 04/13/1994
• Issued: 02/02/1999
• Est. Priority Date: 05/04/1990
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Vickers, Timothy A., Bruice, Thomas W., Ecker, David
• Primary Examiner(s): Guzo, David
• Application Number:
  US08/227,180
• Time in Patent Office: 1,756 Days
• Field of Search: 536/23.1, 536/24.1, 536/24.5, 435/5, 435/6
• US Class Current: 536/24.5
• CPC Class Codes: C12N 15/1132 against retroviridae, e.g. HIV
• CPC Class Codes: C12N 15/63 Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression
• CPC Class Codes: C12N 2310/15 Nucleic acids forming more than 2 strands, e.g. TFOs
• CPC Class Codes: C12N 2310/315 Phosphorothioates

Abstract

Methods for modulating the expression of viral genes are provided by selecting a portion of RNA coded by the gene, said RNA portion having subportions forming a secondary structure, and contacting the RNA with oligonucleotide of 6 to 50 which can bind with at least one of said subportions of the RNA. In accordance with the preferred embodiments, oligonucleotides are designed to bind to RNA secondary structures which are of significance to the expression of the gene coding for said RNA. In accordance with a preferred embodiment, methods of treatment of human immunodeficiency virus are similarly disclosed wherein the oligonucleotides are targeted at the CAR or gag-pol region of HIV RNA.

Claims

1. An oligonucleotide having 6 to 50 bases which binds to at least a portion of the gag-pol region of HIV RNA having secondary structure, said oligonucleotide modulating HIV gag-pol gene expression.

2. The oligonucleotide of claim 1 comprising SEQ. ID. NO. 6, SEQ. ID. NO. 7, SEQ. ID. NO. 8, SEQ. ID. NO. 9 or SEQ. ID. NO. 10.

3. The oligonucleotide of claim 1 comprising the sequence CCC UUC CNN UC (nucleotides 1-11 of SEQ ID NO:

23).

4. The oligonucleotide of claim 1 comprising SEQ. ID. NO. 11, SEQ. ID. NO. 12, SEQ. ID. NO. 13, SEQ. ID. NO. 14, SEQ. ID. NO. 15, SEQ. ID. NO. 16, SEQ. ID. NO. 17 or SEQ. ID. NO. 18.

5. The oligonucleotide of claim 1 comprising SEQ. ID. NO. 23 or SEQ. ID. NO. 24.

6. The oligonucleotide of claim 1 which forms a duplex structure with said region wherein formation of said duplex structure modulates HIV gag-pol gene expression.

7. The oligonucleotide of claim 1 which forms a triplex structure with said region wherin formation of said triplex structure modulates HIV gag-pol gene expression.