Duplex oligonucleotide complexes and methods for gene silencing by RNA interference

Inventors

Yamada, Christina • Khvorova, Anastasia • Kaiser, Rob • Anderson, Emily • Leake, Devin

Assignees

Dharmacon Inc

Abstract

Provided herein are duplex oligonucleotide complexes which can be administered to a cell, tissue or organism to silence a target gene without the aid of a transfection reagent(s). The duplex oligonucleotide complexes of the disclosure include a conjugate moiety that facilitates delivery to a cell, tissue or organism.

Core Innovation

The invention relates to a method for inhibiting expression of a target gene in a cell by delivering a duplex oligonucleotide complex. The complex includes a sense strand and an antisense strand of defined lengths, with defined nucleotide modifications and duplex architecture. The sense strand has a 3′ end and is nineteen nucleotides in length, with nucleotides 1 and 2 and all C and all U nucleotides on the sense strand being 2′O-methyl modified, and the antisense strand is twenty-one nucleotides in length and includes 2′F modified C and U nucleotides.

The duplex is defined by a specific mismatch position and a 3′ end overhang. One nucleotide is mismatched with the sense strand at a defined position within the antisense strand, with the remaining nucleotides complementary to the sense strand. The sense strand and antisense strand form a duplex having a two nucleotide overhang at the 3′ end of the antisense strand, and a phosphate group is present at the 5′ end of the antisense strand.

A cholesterol molecule is attached to the 3′ end of the sense strand via a C5 linker molecule. The disclosed cholesterol conjugation uses a defined cholesterol-linker-sense strand structure, and following delivery the expression of a target protein is decreased.

Claims Coverage

The independent claims include two separate methods for inhibiting expression of a target gene in a cell. Across the independent claims, the inventive features are centered on defined strand lengths, specific 2′ modifications, a single mismatch at a defined antisense position, a two-nucleotide 3′ overhang, cholesterol conjugated to the sense 3′ end via a C5 linker, and a 5′ phosphate on the antisense strand, with decreased target protein expression following delivery.

The independent claims cover delivery of a defined-length sense/antisense duplex with specified 2′ nucleotide modifications, a single mismatch at a defined antisense position, a two-nucleotide 3′ overhang, cholesterol conjugated to the sense 3′ end via a C5 linker, and a 5′ phosphate on the antisense, resulting in decreased expression of a target protein.

Stated Advantages

Documented Applications