MicroRNA vectors
View Patent ↗The present invention relates to gene silencing, and in particular to compositions of microRNA sequences and vectors and to methods of synthesizing such in vitro and in vivo, and to methods of using such to regulate gene expression.
1. A composition comprising a vector, said vector comprising a sequence encoding at least a portion of a BIC miRNA precursor molecule operably linked to a promoter, wherein said portion comprises at least nucleotides 1-111 of SEQ ID NO:47, and wherein said promoter permits expression of said miRNA in a cell.
2. The composition of claim 1 , wherein said portion comprises SEQ ID NO: 49.
3. The composition of claim 1 , wherein said miRNA is produced from said DNA molecule as a hairpin RNA.
4. The composition of claim 1 , wherein said miRNA includes at least a portion of a miR155 miRNA.
5. A composition comprising a vector, said vector comprising a sequence encoding at least a portion of a BIC miRNA precursor molecule operably linked to a promoter, wherein said portion comprises SEQ ID NO:47, wherein approximately bases 29-50 or 68-87 of SEQ ID NO:47 are replaced with sequences complementary to a target mRNA, and wherein said bases 29-50 are at least partially complementary to said bases 68-87.
6. The composition of claim 5 , wherein said miRNA is produced from said DNA molecule as a hairpin RNA.
7. The composition of claim 5 , wherein said miRNA includes at least a portion of a miR155 miRNA.
8. A method for inhibiting the function of a target RNA molecule, comprising transfecting a cell with a DNA molecule comprising a sequence encoding at least a portion of a BIC miRNA precursor molecule operably linked to a promoter, wherein said portion comprises at least nucleotides 1-111 of SEQ ID NO:47, and wherein the promoter can be expressed in the cell, wherein said BIC miRNA encodes an miRNA complementary to a portion of said target RNA molecule.
9. The method of claim 8 , wherein said portion comprises SEQ ID NO: 49.
10. The method of claim 8 , wherein said miRNA is produced from said DNA molecule as a hairpin RNA.
11. The method of claim 8 , wherein said miRNA includes at least a portion of a miR155 miRNA.
12. The method of claim 8 , wherein said cell is in an organism.
13. A method for inhibiting the function of a target RNA molecule, comprising transfecting a cell with a DNA molecule comprising a sequence encoding at least a portion of a BIC miRNA precursor molecule operably linked to a promoter, wherein said portion comprises SEQ ID NO:47, wherein approximately bases 29-50 or 68-87 of SEQ ID NO:47 are replaced with sequences complementary to said target mRNA, and wherein said bases 29-50 are at least partially complementary to said bases 68-87.
14. The method of claim 13 , wherein said miRNA is produced from said DNA molecule as a hairpin RNA.
15. The method of claim 13 , wherein said miRNA includes at least a portion of a miR155 miRNA.
16. The method of claim 13 , wherein said cell is in an organism.