Methods and compositions for regulating RNA import into mitochondria

Patent metadata

Field	Value
Publication number	US-9238041-B2
Application number	US-201213413842-A
Country	US
Kind code	B2
Filing date	Mar 7, 2012
Priority date	May 3, 2011
Publication date	Jan 19, 2016
Grant date	Jan 19, 2016

Abstract

Disclosed herein are methods of manipulating the processing, targeting, and/or internalization of a nucleic acid molecule. Also disclosed are recombinant and/or isolated nucleic acid molecules having a first nucleic acid sequence, which is a wild-type sequence or an altered sequence, directly or indirectly linked to a second nucleic acid sequence which is a mitochondria localization sequence, an RNA import sequence, or a combination thereof, and methods of using thereof.

First claim

We claim:

1. A method of manipulating the processing, targeting, and/or internalization of a nucleic acid molecule which comprises one or more of the following steps
providing an altered tRNA aminoacyl stem that is directly or indirectly linked to the nucleic acid molecule to thereby enable the nucleic acid molecule to escape the nucleus of a cell; providing a mitochondria localization sequence that is directly or indirectly linked to the nucleic acid molecule to thereby cause the nucleic acid molecule to localize in the proximity of a mitochondrion; and
providing an RNA import sequence that is directly or indirectly linked to the nucleic acid molecule to thereby cause the nucleic acid molecule to be internalized by a mitochondrion, and
wherein the RNA import sequence is selected from the group consisting of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:38, SEQ ID NO:39, sequences which are about 15-30 nucleotides long and comprise SEQ ID NO:39, and sequences complementary thereto.

2. The method of claim 1 , wherein the nucleic acid molecule is a nucleus-encoded non-coding RNA.

3. The method of claim 1 , wherein the nucleic acid molecule is a wild-type sequence of a gene a fragment thereof or its complement in the mitochondrial genome of a mammalian subject.

4. The method of claim 1 , where the nucleic acid molecule is a RNA:DNA hybrid.

5. The method of claim 1 ,
wherein the sequences are capable of forming a single stem-loop.

6. The method of claim 1 , wherein the mitochondrial localization sequence is mammalian.

7. The method of claim 1 , wherein the mitochondrial localization sequence is SEQ ID NO:34 or its complement.

8. The method of claim 1 , wherein the nucleic acid sequence is not natively associated with the mitochondria localization sequence or the RNA import sequence.

9. The method of claim 1 , wherein the nucleic acid sequence is RNA or ssDNA.

10. The method of claim 1 , wherein the nucleic acid molecule is a microRNA.

11. The method of claim 1 , wherein the nucleic acid molecule is a riboenzyme.

Assignees

• Teitell Michael A
• Koehler Carla M
• Wang Geng
• Univ California

Inventors

• Teitell Michael A
• Koehler Carla M
• Wang Geng

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• C12N15/1137

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title

• C12Y207/07008

  Polyribonucleotide nucleotidyltransferase (2.7.7.8), i.e. polynucleotide phosphorylase · CPC title

• C12N15/87

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title

• A61K31/7088 Primary

  Compounds having three or more nucleosides or nucleotides · CPC title