Compositions and methods for transfection of RNA and controlled stabilization of transfected RNA
The present invention provides reagents and methods for RNA transfection and protein expression.
1. A recombinant nucleic acid, comprising
(a) an RNA molecule; and
(b) a first recombinant RNA comprising two or more copies of a 3′ untranslated region (UTR) of an RNA virus selected from the group consisting of Venezuelan equine encephalitis virus (VEEV) and Sinbis virus;
wherein the first recombinant RNA is coupled to the 3′ end of the RNA molecule.
2. The recombinant nucleic acid of claim 1 , wherein the first recombinant RNA comprises three or more copies of the 3′ UTR.
3. The recombinant nucleic acid of claim 1 , wherein the first recombinant RNA comprises six or more copies of the 3′ UTR.
4. The recombinant nucleic acid of claim 1 , wherein the recombinant RNA comprises an RNA transcript encoded by SEQ ID NO:1.
5. An isolated host cell transfected with the recombinant nucleic acid of claim 1 .
6. An isolated host cell transfected with the recombinant nucleic acid of claim 4 .
7. A method for RNA transfection, comprising transfecting the recombinant nucleic acid of claim 1 into a host cell.
8. A method for RNA transfection, comprising transfecting the recombinant nucleic acid of claim 4 into a host cell.
9. The recombinant nucleic acid of claim 1 , wherein the RNA molecule is selected from the group consisting of an mRNA molecule and an siRNA molecule.
10. An isolated host cell transfected with the recombinant nucleic acid of claim 3 .
11. A method for RNA transfection, comprising transfecting the recombinant nucleic acid of claim 3 into a host cell.
12. The recombinant nucleic acid of claim 3 , wherein the RNA molecule is selected from the group consisting of an mRNA molecule and an siRNA molecule.
13. The recombinant nucleic acid of claim 4 , wherein the RNA molecule is selected from the group consisting of an mRNA molecule and an siRNA molecule.
14. The recombinant nucleic acid of claim 4 , wherein the first recombinant RNA comprises six or more copies the 3′ UTR.
15. The recombinant nucleic acid of claim 1 , wherein the recombinant RNA comprises an RNA transcript encoded by SEQ ID NO:2.
16. The recombinant nucleic acid of claim 15 , wherein the first recombinant RNA comprises six or more copies the 3′ UTR.
17. An isolated host cell transfected with the recombinant nucleic acid of claim 15 .
18. A method for RNA transfection, comprising transfecting the recombinant nucleic acid of claim 15 into a host cell.
19. An isolated host cell transfected with the recombinant nucleic acid of claim 16 .
20. A method for RNA transfection, comprising transfecting the recombinant nucleic acid of claim 16 into a host cell.