Methods and compositions for the production of siRNAs
The technology described herein relates to siRNAs, e.g., methods and compositions relating to the production of siRNAs in bacterial cells.
1. A bacterial cell comprising a viral siRNA-binding polypeptide and a dsRNA comprising an exogenous nucleic acid sequence substantially complementary to a target RNA.
2. The bacterial cell of claim 1 , wherein the siRNA-binding polypeptide comprises a purification tag.
3. The bacterial cell of claim 1 , wherein the siRNA-binding polypeptide is encoded by a nucleic acid.
4. The bacterial cell of claim 1 , wherein the siRNA-binding polypeptide is selected from the group consisting of:
p19 polypeptide; tombusvirus p19 polypeptide; B2 polypeptide; HC-Pro polypeptide; p38 polypeptide; p122 polypeptide; p130 polypeptide; p21 polypeptide; p1b polypeptide; and NS3 polypeptide.
5. The bacterial cell of claim 1 , wherein the dsRNA is greater than 21 nucleotides in length.
6. The bacterial cell of claim 1 , wherein the dsRNA is a hairpin RNA.
7. The bacterial cell of claim 1 , wherein the bacterial cell expresses an RNase III polypeptide.
8. The bacterial cell of claim 1 , wherein the bacterial cell expresses an RNase III polypeptide encoded by an exogenous nucleic acid sequence.
9. The bacterial cell of claim 1 , wherein the bacterial cell is an Escherichia coli cell.
10. The bacterial cell of claim 1 , wherein at least one of the siRNA-binding polypeptide and the dsRNA are constitutively expressed.
11. The bacterial cell of claim 1 , wherein at least one of the siRNA-binding polypeptide and the dsRNA are inducibly expressed.
12. The bacterial cell of claim 1 , wherein the DNA encoding at least one of the siRNA-binding polypeptide or the dsRNA is part of a plasmid.
13. The bacterial cell of claim 1 , wherein the dsRNA comprises nucleic acid sequences substantially complementary to a multiplicity of target RNAs.