Methods and compositions for prevention or treatment of a disease
Methods and compositions for treatment of a genetic disease are provided.
1. A fusion molecule comprising (i) an engineered nuclease that binds to a target site within SEQ NOs:3, 4, 5 or 6 of an endogenous gene that produces a microRNA (miRNA), and (ii) a cleavage domain.
2. A polynucleotide encoding one or more fusion molecules of claim 1 .
3. An isolated cell comprising one or more fusion molecules according to claim 1 .
4. The cell of claim 3 , wherein the cell is a stem cell.
5. The cell of claim 3 , wherein the stem cell is selected from the group consisting of an embryonic stem cell (ESC), an induced pluripotent stem cell (iPSC), a hepatic stem cell and a liver stem cell.
6. The fusion molecule of claim 1 , wherein the engineered nuclease is a zinc finger nuclease.
7. The zinc finger nuclease of claim 6 wherein the endogenous gene is MiR-34 and the zinc finger protein comprises 5 zinc, finger domains, each domain comprising a recognition helix region, wherein the zinc finger protein comprises the recognition helix regions as shown in a single row of Table III.
8. A kit comprising a fusion molecule according to claim 1 .
9. A kit comprising a polynucleotide according to claim 2 .
10. A method of cleaving an endogenous gene that produces an miRNA in a cell, the method comprising:
introducing, into the cell, one or more expression vectors comprising at least one polynucleotide according to claim 2 , under conditions such that the one or more fusion molecules are expressed and the miRNA-producing gene is cleaved.
11. The method of claim 10 , wherein the polynucleotide comprises an AAV vector.
12. The method of claim 10 , wherein the polynucleotide is an mRNA.