Rna sequence-specific mediators of rna interference

Patent metadata

Field	Value
Publication number	US-2020270602-A1
Application number	US-201916580016-A
Country	US
Kind code	A1
Filing date	Sep 24, 2019
Priority date	Mar 30, 2000
Publication date	Aug 27, 2020
Grant date	—

Abstract

The present invention relates to a Drosophila in vitro system which was used to demonstrate that dsRNA is processed to RNA segments 21-23 nucleotides (nt) in length. Furthermore, when these 21-23 nt fragments are purified and added back to Drosophila extracts, they mediate RNA interference in the absence of long dsRNA, Thus, these 21-23 nt fragments are the sequence-specific mediators of RNA degradation. A molecular signal, which may be their specific length, must be present in these 21-23 nt fragments to recruit cellular factors involved in RNAi. This present invention encompasses these 21-23 nt fragments and their use for specifically inactivating gene function. The use of these fragments (or chemically synthesized oligonucleotides of the same or similar nature) enables the targeting of specific mRNAs for degradation in mammalian cells, where the use of long dsRNAs to elicit RNAi is usually not practical, presumably because of the deleterious effects of the interferon response. This specific targeting of a particular gene function is useful in functional genomic and therapeutic applications.

First claim

1 . Isolated RNA of from about 21 to about 23 nucleotides that mediates RNA interference of an mRNA to which it corresponds.

2 . Isolated RNA of claim 1 that comprises a terminal 3′ hydroxyl group.

3 . Isolated RNA of claim 1 which is chemically synthesized RNA or an analog of a naturally occurring RNA.

4 . An analog of isolated RNA of claim 1 , wherein the analog differs from the RNA of claim 1 by the addition, deletion, substitution or alteration of one or more nucleotides.

5 .- 16 . (canceled)

17 . A method of mediating RNA interference of mRNA of a gene in a cell or organism comprising:
(a) introducing RNA of from about 21 to about 23 nucleotides which targets the mRNA of the gene for degradation into the cell or organism; (b) maintaining the cell or organism produced in (a) under conditions under which degradation of the mRNA occurs, thereby mediating RNA interference of the mRNA of the gene in the cell or organism.

18 . The method of claim 17 wherein the RNA of (a) is a chemically synthesized RNA or an analog of naturally occurring RNA.

19 . The method of claim 17 , wherein the gene encodes a cellular mRNA or a viral mRNA.

20 .- 35 . (canceled)

36 . A method of treating a disease or condition associated with the presence of a protein in an individual comprising administering to the individual RNA of from about 21 to about 23 nucleotides that targets the mRNA of the protein for degradation.

37 . The method of claim 36 wherein RNA of from about 21 to about 23 nucleotides is chemically synthesized or an analog of RNA that mediates RNA interference.

38 .- 42 . (canceled)

43 . A pharmaceutical composition comprising RNA of from about 21 to about 23 nucleotides that mediates RNA interference and an appropriate carrier.

44 .- 50 . (canceled)

Assignees

• Max-Planck-Gesellschaft Zur Fõrderung Der Wss Ev
• Massachusetts Inst Technology
• Whitehead Inst Biomedical Res
• Univ Massachusetts

Inventors

• Tuschl Thomas
• Zamore Phillip D
• Sharp Phillip A
• Bartel David P

Classifications

• A01K67/64

  Genetically modified nematodes · CPC title

• C12N15/111

  General methods applicable to biologically active non-coding nucleic acids · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• C12N15/1079 Primary

  Screening libraries by altering the phenotype or phenotypic trait of the host (reporter assays C12N15/1086) · CPC title

• C12N2310/321

  2'-O-R Modification · CPC title