Multimeric coding nucleic acid and uses thereof
View Patent ↗The present invention provides, among other things, multimeric coding nucleic acids that exhibit superior stability for in vivo and in vitro use. In some embodiments, a multimeric coding nucleic acid (MCNA) comprises two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends.
1. A method of delivering a multimeric coding nucleic acid (MCNA) for in vivo protein production, comprising administering to a subject in need of the MCNA, wherein the MCNA comprises two messenger RNAs (mRNAs) linked at 3′ ends, via a stable linkage, such that the multimeric coding nucleic acid has two 5′ ends, and wherein the stable linkage is an oligonucleotide bridge comprising an internal 3′-to-3′ inverted phosphodiester linkage.
2. The method of claim 1 , wherein each of the mRNAs encodes a protein of interest.
3. The method of claim 2 , wherein each of the mRNAs encodes a same protein.
4. The method of claim 2 , wherein each of the mRNAs encodes a distinct protein.
5. The method of claim 1 , wherein the mRNAs comprise a 3′ UTR.
6. The method of claim 5 , wherein the 3′ UTR comprises a plurality of multi-A segments with spacers in between.
7. The method of claim 1 , wherein the oligonucleotide bridge comprises nucleosides selected from the group consisting of 2′-OMe-A, 2′-OMe-G, 2′-OMe-C, 2′-OMe-U, 2′-F-A, 2′-F-G, 2′-F-C, 2′-F-U, LNA-A, LNA-G, LNA-C, LNA-U, N6-methyl-adenosine, 2-thiouridine (2sU), 5-methyl-cytidine (5mC), pseudouridine (ΨU), and 1-methyl-pseudouridine.
8. The method of claim 1 , wherein the mRNAs comprise one or more modified nucleosides.
9. The method of claim 8 , wherein the modified nucleosides are selected from the group consisting of 2′-OMe-A, 2′-OMe-G, 2′-OMe-C, 2′-OMe-U, 2′-F-A, 2′-F-G, 2′-F-C, 2′-F-U, LNA-A, LNA-G, LNA-C, LNA-U, N6-methyl-adenosine, 2-thiouridine (2sU), 5-methyl-cytidine (5mC), pseudouridine (ΨU), and 1-methyl-pseudouridine.
10. The method of claim 1 , wherein each of the mRNAs encodes an enzyme, a receptor, a ligand, a light chain or heavy chain of an antibody, a nuclease, or a DNA-binding protein.
11. The method of claim 1 , wherein the MCNA is encapsulated or complexed with a delivery vehicle.
12. The method of claim 11 , wherein the delivery vehicle is selected from the group consisting of liposomes, lipid nanoparticles, solid-lipid nanoparticles, polymers, viruses, sol-gels, and nanogels.
13. The method of claim 5 , wherein the 3′ UTR does not include a polyA tail.
14. The method of claim 1 , wherein each of the mRNAs is unmodified.
15. The method of claim 1 , wherein the oligonucleotide bridge is unmodified.
16. The method of claim 1 , wherein each of the mRNAs has a 5′ cap.