Production of stable non-polyadenylated RNAs
The invention relates in aspects to hybrid RNAs lacking a poly-A tail and nucleic acid vectors for expressing the RNA. The hybrid RNAs in some instances have a 3′ terminal stabilizing triple helical structure. Related methods for expressing said RNAs in vivo and in vitro are also disclosed.
1. A hybrid nucleic acid comprising:
an RNA molecule lacking a poly-A tail, linked to a heterologous RNA stabilizing terminal sequence, wherein the heterologous RNA stabilizing terminal sequence has a triple helix conformation.
2. The hybrid nucleic acid of claim 1 , wherein the RNA molecule is a cytoplasmic RNA or a nuclear RNA.
3. The hybrid nucleic acid of claim 1 , wherein the RNA molecule is a mRNA which lacks a poly-A tail.
4. The hybrid nucleic acid of claim 1 , wherein the RNA molecule is a noncoding RNA.
5. The hybrid nucleic acid of claim 1 , wherein the RNA molecule is a eukaryotic RNA.
6. The hybrid nucleic acid of claim 1 , wherein the RNA molecule is a mammalian RNA.
7. The hybrid nucleic acid of claim 1 , wherein the RNA molecule is a plant RNA.
8. The hybrid nucleic acid of claim 1 , wherein the RNA molecule is a human RNA.
9. A hybrid nucleic acid comprising:
an RNA molecule lacking a poly-A tail, linked to a heterologous RNA stabilizing terminal sequence, wherein the heterologous RNA stabilizing terminal sequence is a MALAT1 terminal sequence.
10. The hybrid nucleic acid of claim 1 , wherein the heterologous RNA stabilizing terminal sequence is a MEN β terminal sequence.
11. The hybrid nucleic acid of claim 1 , wherein the heterologous RNA stabilizing terminal sequence is a U-rich sequence.
12. The hybrid nucleic acid of claim 1 , wherein the heterologous RNA stabilizing terminal sequence is an A-rich sequence.
13. The hybrid nucleic acid of claim 1 , wherein the heterologous RNA stabilizing terminal sequence is a U-rich and A-rich sequence.
14. The hybrid nucleic acid of claim 1 , wherein the heterologous RNA stabilizing terminal sequence is a C-rich and G-rich sequence.
15. The hybrid nucleic acid of claim 1 , wherein the RNA molecule corresponds to a reporter molecule.
16. A vector comprising: a nucleic acid encoding an RNA molecule, a promoter upstream of the nucleic acid encoding the RNA molecule and a terminal nucleic acid sequence downstream of the nucleic acid encoding the RNA molecule, wherein the vector includes a nucleic acid sequence that produces the hybrid nucleic acid of claim 1 .
17. The hybrid nucleic acid of claim 1 , wherein nucleic acid includes at least one chemical or natural modification.
18. A method for enhancing translation of an RNA, comprising,
expressing in a cell an isolated cytoplasmic RNA lacking a poly A tail, wherein the cytoplasmic RNA has a 3′ terminal sequence effective for enhancing translation of the RNA in the cell, wherein the cytoplasmic RNA is the hybrid nucleic acid of claim 1 .
19. A method for expressing an RNA lacking a poly-A tail, comprising:
expressing in a cell an isolated nucleic acid comprising an RNA having a 3′ heterologous terminal sequence and lacking a poly A tail, wherein the RNA is the hybrid nucleic acid of claim 1 .
20. A composition comprising:
an RNA molecule, lacking a poly-A tail, linked to a heterologous RNA stabilizing terminal sequence, formulated in a nanoparticle, wherein the heterologous RNA stabilizing terminal sequence has a triple helix conformation.
21. The composition of claim 20 , wherein RNA includes at least one chemical or natural modification.