Compositions and methods for delivery of nucleic acids to cells
Compositions and methods of use thereof for delivering nucleic acid cargo into cells are provided. The compositions typically include (a) a 3E10 monoclonal antibody or an antigen binding, cell-penetrating fragment thereof; a monovalent, divalent, or multivalent single chain variable fragment (scFv); or a diabody; or humanized form or variant thereof, and (b) a nucleic acid cargo including, for example, a nucleic acid encoding a polypeptide, a functional nucleic acid, a nucleic acid encoding a functional nucleic acid, or a combination thereof. Elements (a) and (b) are typically non-covalently linked to form a complex.
1. A composition comprising a non-covalent complex of (a) an antibody or antigen-binding fragment thereof, and (b) an mRNA,
wherein the antibody or antigen-binding fragment thereof comprises heavy chain variable region (V H ) complementarity determining regions (CDRs) having the amino acid sequences of SEQ ID NO:16, SEQ ID NO:17, and SEQ ID NO:18, and light chain variable region (V L ) CDRs having the amino acid sequences of SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26.
2. The composition according to claim 1 , wherein the antibody or antigen-binding fragment thereof comprises a V H having an amino acid sequence that is at least 95% identical to SEQ ID NO:2 and a V L having an amino acid sequence that is at least 95% identical to SEQ ID NO:7 or 8.
3. The composition according to claim 2 , wherein the V H has the amino acid sequence of SEQ ID NO:2 and the V L has the amino acid sequence of SEQ ID NO:7 or 8.
4. The composition according to claim 1 , wherein the antibody or antigen-binding fragment thereof comprises a V H having an amino acid sequence that is at least 95% identical to SEQ ID NO: 6 and a V L having an amino acid sequence that is at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 11 and 53-55.
5. The composition according to claim 4 , wherein the V H has an amino acid sequence of SEQ ID NO: 6 and the V L has an amino acid sequence selected from the group consisting of SEQ ID NOs: 11 and 53-55.
6. The composition according to claim 1 , wherein the antibody or antigen-binding fragment thereof is a humanized antibody.
7. The composition according to claim 1 , wherein the antibody or antigen-binding fragment thereof is a monovalent single chain variable fragment (scFv), divalent scFv, or multivalent scFv.
8. The composition of claim 1 , wherein the antibody or antigen-binding fragment thereof is a bispecific antibody.
9. The composition of claim 8 , wherein the bispecific antibody comprises:
a first heavy chain or antigen-binding fragment thereof comprising CDRs having the amino acid sequences of SEQ ID NO:16, SEQ ID NO:17, and SEQ ID NO:18, and a first light chain or antigen binding fragment thereof comprising CDRs having the amino acid sequences of SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26; and
a second heavy chain, or antigen-binding fragment thereof, and a second light chain, or antigen binding fragment thereof, that associate to specifically bind a target cell-type, tissue, or organ.
10. The composition according to claim 1 , wherein the mRNA encodes a polypeptide ligand for a receptor of an immune cell.
11. The composition according to claim 10 , wherein the polypeptide ligand is a cytokine.
12. The composition according to claim 10 , wherein the polypeptide ligand stimulates the immune system of the subject.
13. The composition according to claim 1 , wherein the mRNA encodes a polypeptide that is defective in a genetic disease.
14. The composition according to claim 13 , wherein the mRNA encodes dystrophin.
15. The composition according to claim 13 , wherein the mRNA encodes utrophin.
16. The composition according to claim 1 , wherein the mRNA encodes an antigen.
17. The composition according to claim 16 , wherein the antigen is a viral antigen.
18. The composition according to claim 17 , wherein the viral antigen is a SARS-CoV-2 antigen.
19. The composition according to claim 1 , wherein the mRNA encodes a pro-apoptotic factor.
20. The composition according to claim 19 , wherein the pro-apoptotic factor is a BH3-only pro-apoptotic protein.
21. The composition according to claim 1 , wherein the mRNA encodes a tumor suppressor.
22. The composition according to claim 1 , further comprising a pharmaceutically acceptable excipient.