Engineered nucleic acids and methods of use thereof

1. A pharmaceutical composition comprising:

a lipid-based nanoparticle comprising a synthetic messenger ribonucleic acid (mRNA) encoding a defensin polypeptide in an amount effective to permit production of the defensin polypeptide in a cell, wherein the synthetic mRNA comprises a translatable region that contains at least one nucleoside modification, and wherein 75-100% of uridine nucleotides in the synthetic mRNA are modified.

2. The pharmaceutical composition of claim 1 , wherein the at least one nucleoside modification is selected from the group consisting of pyridin-4-one ribonucleoside, 5-aza-uridine, 2-thio-5-aza-uridine, 2-thiouridine, 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxyuridine, 3-methyluridine, 5-carboxymethyl-uridine, 1-carboxymethyl-pseudouridine, 5-propynyl-uridine, 1-propynyl-pseudouridine, 5-taurinomethyluridine, 1-taurinomethyl-pseudouridine, 5-taurinomethyl-2-thio-uridine, 1-taurinomethyl-4-thio-uridine, 5-methyl-uridine, 1-methyl-pseudouridine, 4-thio-1-methyl-pseudouridine, 2-thio- 1 -methyl-pseudouridine, 1-methyl- 1 -deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine, dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2-methoxyuridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, 5-aza-cytidine, pseudoisocytidine, 3-methyl-cytidine, N4-acetylcytidine, 5-formylcytidine, N4-methylcytidine, 5-hydroxymethylcytidine, 1-methyl-pseudoisocytidine, pyrrolo-cytidine, pyrrolo-pseudoisocytidine, 2-thio-cytidine, 2-thio-5-methyl-cytidine, 4-thio-pseudoisocytidine, 4-thio-1-methyl-pseudoisocytidine, 4-thio-1-methyl-1-deaza-pseudoisocytidine, 1-methyl-1-deaza-pseudoisocytidine, zebularine, 5-aza-zebularine, 5-methyl-zebularine, 5-aza-2-thio-zebularine, 2-thio-zebularine, 2-methoxy-cytidine, 2-methoxy-5-methyl-cytidine, 4-methoxy-pseudoisocytidine, 4-methoxy-1-methyl-pseudoisocytidine, 2-aminopurine, 2, 6-diaminopurine, 7-deaza-adenine, 7-deaza-8-aza-adenine, 7-deaza-2-aminopurine, 7-deaza-8-aza-2-aminopurine, 7-deaza-2,6-diaminopurine, 7-deaza-8-aza-2,6-diaminopurine, 1-methyladenosine, N6-methyladenosine, N6-isopentenyladenosine, N6-(cis-hydroxyisopentenyl)adenosine, 2-methylthio-N6-(cis-hydroxyisopentenyl) adenosine, N6-glycinylcarbamoyladenosine, N6-threonylcarbamoyladenosine, 2-methylthio-N6-threonyl carbamoyladenosine, N6,N6-dimethyladenosine, 7-methyladenine, 2-methylthio-adenine, 2-methoxy-adenine, inosine, 1-methyl-inosine, wyosine, wybutosine, 7-deaza-guanosine, 7-deaza-8-aza-guanosine, 6-thio-guanosine, 6-thio-7-deaza-guanosine, 6-thio-7-deaza-8-aza-guanosine, 7-methyl-guanosine, 6-thio-7-methyl-guanosine, 7-methylinosine, 6-methoxy-guanosine, 1-methylguanosine, N2-methylguanosine, N2,N2-dimethylguanosine, 8-oxo-guanosine, 7-methyl-8-oxo-guanosine, 1-methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine, and N2,N2-dimethyl-6-thio-guanosine.

3. The pharmaceutical composition of claim 1 , wherein the composition is formulated for administration via a route selected from the group consisting of: systemic, local, intravenous, topical, oral, administration via a dressing, administration via a bandage, rectal, vaginal, intramuscular, transarterial, intraperitoneal, intranasal, subcutaneous, endoscopic, transdermal and intrathecal.

4. The pharmaceutical composition of claim 3 , wherein the route is intravenous.

5. The pharmaceutical composition of claim 1 , wherein the defensin polypeptide is selected from the group consisting of α-defensins, and β-defensins and θ-defensins.

6. The pharmaceutical composition of claim 1 , wherein the anti-microbial polypeptide is hBD-2 (SEQ ID NO: 191 or 192).

7. A kit comprising:

(a) a synthetic messenger ribonucleic acid (mRNA) encoding a defensin polypeptide packaged in a container, wherein the synthetic mRNA comprises a translatable region that contains at least one nucleoside modification, and wherein 75-100% of uridine nucleotides in the synthetic mRNA are modified; and

(b) a pharmaceutically acceptable carrier packaged in a container.

8. The pharmaceutical composition of claim 5 , wherein the defensin is an α-defensin selected from the group consisting of neutrophil defensin 1, defensin alpha 1, neutrophil defensin 3, neutrophil defensin 4, defensin 5 and defensin 6.

9. The pharmaceutical composition of claim 5 , wherein the defensin is a β-defensin selected from the group consisting of beta-defensin 1, beta-defensin 2, beta-defensin 103, beta-defensin 107, beta-defensin 110 and beta-defensin 136.

10. The pharmaceutical composition of claim 5 , wherein the at least one nucleoside modification is pseudouridine.

11. The pharmaceutical composition of claim 5 , wherein 100% of uridine nucleotides in the synthetic mRNA are modified.

12. The pharmaceutical composition of claim 5 , wherein the synthetic mRNA further comprises a 5′ untranslated region that contains at least one nucleoside modification.

13. The pharmaceutical composition of claim 5 , wherein the synthetic mRNA further comprises a 5′ untranslated region that contains at least two nucleoside modifications.

14. The pharmaceutical composition of claim 5 , wherein the synthetic mRNA further comprises a 3′ untranslated region that contains at least one nucleoside modification.

15. The pharmaceutical composition of claim 5 , wherein the synthetic mRNA further comprises a 3′ untranslated region that contains at least two nucleoside modifications.

16. A lipid-based nanoparticle comprising a synthetic messenger ribonucleic acid (mRNA) encoding a defensin polypeptide in an amount effective to permit production of the defensin polypeptide in a cell,

wherein 75-100% of uridine nucleotides in the synthetic mRNA are modified, and

wherein the synthetic mRNA comprises a translatable region, a 5′ untranslated region and a 3′ untranslated region, each of which contains at least one nucleoside modification.

17. The lipid-based nanoparticle of claim 16 , wherein the at least one nucleoside modification is pseudouridine.