RNA-mediated gene modulation

• US 8,609,831 B2
• Filed: 09/16/2003
• Issued: 12/17/2013
• Est. Priority Date: 09/16/2002
• Status: Expired due to Fees

• Current Assignee: University of Southern California
• Original Assignee: University of Southern California
• Inventors: Lin, Shi-Lung, Ying, Shao-Yao
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US10/663,875
• Publication Number:
  US 20040253604A1
• Time in Patent Office: 3,745 Days
• Field of Search: None
• US Class Current: 536/24.5
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/30 Chemical structure

Abstract

An isolated RNA comprising an intron RNA that is released in a cell, thereby modulating the function of a target gene. Also disclosed are a composition comprising a chemokine and an isolated RNA of the invention or a DNA template for the isolated RNA, a composition comprising one or more agents that induce RNA-mediated modulation of the functions of two or more target genes in a cell, and methods of using these compositions for modulating the functions of genes in a cell.

Claims

1. An isolated RNA comprising:

an artificial intron comprising a splice donor site, a splice acceptor site, a branch site, a poly-pyrimidine tract and at least one aberrant RNA selected from the group consisting of a short-temporary RNA, small-interfering RNA, short-hairpin RNA, long deoxyribonucleotide-containing RNA and ribozyme RNA,

wherein said aberrant RNA is an RNA capable of silencing a target gene when released in a cell.

2. The isolated RNA of claim 1, wherein the cell is a mammalian cell.

3. The isolated RNA of claim 1, wherein the splice donor site contains 5′

-AGGUAAGU-3′

.

4. The isolated RNA of claim 1, wherein the splice acceptor site contains 5′

-CCACAGC-3′

.

5. The isolated RNA of claim 1, wherein the branch site contains 5′

-UACUAAC-3.

6. The isolated RNA of claim 1, wherein the cell is a eukaryotic cell.

8. A cultivated cell comprising the isolated RNA of claim 1.

9. A composition comprising the isolated RNA of claim 1.

10. The isolated RNA of claim 1, wherein the artificial intron contains a sequence that targets an exon of the target gene, wherein the target gene is integrin β

1.

11. The isolated RNA of claim 6, wherein the artificial intron contains a sequence that targets an exon of the target gene, wherein the target gene is integrin β

1.

7. An isolated RNA comprising:

an artificial intron RNA comprising at least one aberrant RNA selected from the group consisting of a short-temporary RNA, small-interfering RNA, short-hairpin RNA, long deoxyribonucleotide-containing RNA (D-RNA) and ribozyme RNA, said aberrant RNA being capable of silencing a target gene when released in a eukaryotic cell and wherein the artificial intron RNA further comprises a splice donor site that includes 5′

-AGGUAAGU-3′

, a splice acceptor site that includes 5′

-CCACAGC-3′

, a branch site that includes 5′

-UACUAAC-3′

, a poly-pyrimidine tract that includes 5′

-UUCUUUUUUUC-3′

(SEQ ID NO;

2), or a combination thereof.

12. The isolated RNA of claim 7, wherein the artificial intron contains a sequence that targets an exon of the target gene, wherein the target gene is integrin β

1.