IP Library › Granted Patent US 12,618,065
Granted Patent B2
US 12,618,065 · App. 16/651,814 · Granted May 5, 2026

MiRNA detargeting system for tissue specific interference

Inventor: Scott Quenton Harper (Powell, OH)
Assignee: RESEARCH INSTITUTE AT NATIONWIDE CHILDREN'S HOSPITAL
C12N15/113A61K35/76C12N7/00C12N15/86A61K48/00C12N2750/14143
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Quick Facts
Patent No.
US 12,618,065
App. No.
16/651,814
Granted
May 5, 2026
Kind
B2
Abstract

The present disclosure relates to a tissue-specific promoter system for expressing microRNA (miRNA) for RNA interference-based methods of gene therapy. In these systems, the miRNA will inhibit gene expression or replace natural miRNA expression using microRNA.

Claims (32)

1 . A method of inhibiting expression of the double homeobox 4 (DUX4) gene in a cell comprising contacting the cell with a recombinant adeno-associated virus comprising a nucleic acid comprising

(a) a nucleotide sequence encoding a U6 promoter;

(b) a nucleotide sequence encoding a mature guide strand of a miRNA targeting DUX4 mRNA, wherein the mature guide strand of the miRNA comprises the nucleotide sequence of SEQ ID NO: 7397 or 8147;

(c) a nucleotide sequence comprising at least one copy of a detargeting sequence comprising the mir-122 binding site having the nucleotide sequence of SEQ ID NO: 5 or 66, and/or at least one copy of a detargeting sequence comprising the miR-208 binding site having the nucleotide sequence of SEQ ID NO: 6 or 67; and

(d) 5-6 thymidines at the 5′ end.

2 . A method of delivering double homeobox 4 (DUX4) miRNA-encoding DNA to the skeletal muscle of an animal in need thereof, comprising contacting the skeletal muscle with a recombinant adeno-associated virus comprising a nucleic acid comprising

(a) a nucleotide sequence encoding a U6 promoter;

(b) a nucleotide sequence encoding a mature guide strand of a miRNA targeting DUX4 mRNA, wherein the mature guide strand of the miRNA comprises the nucleotide sequence of SEQ ID NO: 7397 or 8147;

(c) a nucleotide sequence comprising at least one copy of a detargeting sequence comprising the mir-122 binding site having the nucleotide sequence of SEQ ID NO: 5 or 66, and/or at least one copy of a detargeting sequence comprising the miR-208 binding site having the nucleotide sequence of SEQ ID NO: 6 or 67; and

(d) 5-6 thymidines at the 5′ end.

3 . A method of treating facioscapulohumeral muscular dystrophy in a subject comprising administering to the subject an effective amount of a recombinant adeno-associated virus comprising

(a) a nucleotide sequence encoding a U6 promoter;

(b) a nucleotide sequence encoding a mature guide strand of a miRNA targeting DUX4 mRNA, wherein the mature guide strand of the miRNA comprises the nucleotide sequence of SEQ ID NO: 7397 or 8147;

(c) a nucleotide sequence comprising at least one copy of a detargeting sequence comprising the mir-122 binding site having the nucleotide sequence of SEQ ID NO: 5 or 66, and/or at least one copy of a detargeting sequence comprising the miR-208 binding site having the nucleotide sequence of SEQ ID NO: 6 or 67; and

(d) 5-6 thymidines at the 5′ end.

4 . The method of claim 2 , wherein the recombinant adeno-associated virus is administered by intramuscular injection, transdermal transport, injection into the blood stream or injection into the liver.

5 . The method of claim 3 , wherein the recombinant adeno- associated virus is administered by intramuscular injection, transdermal transport, injection into the blood stream or injection into the liver.

6 . The method of claim 1 , wherein the U6 promoter comprises the nucleotide sequence of SEQ ID NO: 3.

7 . The method of claim 1 , wherein the nucleic acid comprises the polynucleotide sequence of any one of SEQ ID NOS: 1, 2, and 10913-10968.

8 . The method of claim 1 , wherein the adeno-associated virus (AAV) is AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13, AAV rh.74 or AAV-B1.

9 . The method of claim 1 , wherein the adeno-associated virus (AAV) is AAV6, AAV rh.74 or AAV-B1.

10 . The method of claim 2 , wherein the U6 promoter comprises the nucleotide sequence of SEQ ID NO: 3.

11 . The method of claim 2 , wherein the nucleic acid comprises the polynucleotide sequence of any one of SEQ ID NOS: 1, 2, and 10913-10968.

12 . The method of claim 2 , wherein the adeno-associated virus (AAV) is AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13, AAV rh.74 or AAV-B1.

13 . The method of claim 2 , wherein the adeno-associated virus (AAV) is AAV6, AAV rh.74 or AAV-B1.

14 . The method of claim 3 , wherein the U6 promoter comprises the nucleotide sequence of SEQ ID NO: 3.

15 . The method of claim 3 , wherein the nucleic acid comprises the polynucleotide sequence of any one of SEQ ID NOS: 1, 2, and 10913-10968.

16 . The method of claim 3 , wherein the adeno-associated virus (AAV) is AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13, AAV rh.74 or AAV-B1.

17 . The method of claim 3 , wherein the adeno-associated virus (AAV) is AAV6, AAV rh.74 or AAV-B1.

18 . The method of claim 1 , wherein the U6 promoter comprises the nucleotide sequence of SEQ ID NO: 4.

19 . The method of claim 2 , wherein the U6 promoter comprises the nucleotide sequence of SEQ ID NO: 4.

20 . The method of claim 3 , wherein the U6 promoter comprises the nucleotide sequence of SEQ ID NO: 4.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 12, 2020
From: HARPER, SCOTT QUENTON
To: RESEARCH INSTITUTE AT NATIONWIDE CHILDREN'S HOSPITAL
Reel/Frame 052926/0730 →
Continuity (2)
Provisional Application 62566966 · Oct 2, 2017
Related Publication 20200248179A1 · Aug 6, 2020
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