Modified polynucleotides for the production of proteins

Abstract

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

1 . An isolated polynucleotide comprising;
 (a) a first region of linked nucleosides, said first region encoding a polypeptide of interest, said polypeptide of interest selected from the group consisting of SEQ ID NOs 8922-17687;   (b) a first flanking region located at the 5′ terminus of said first region comprising;
 (i) a sequence of linked nucleosides selected from the group consisting of the native 5′ UTR of any of the nucleic acids that encode any of SEQ ID NOs 8922-17687, SEQ ID NOs: 1-4 and functional variants thereof; and 
 (ii) at least one 5′ terminal cap; 
   (c) a second flanking region located at the 3′ terminus of said first region comprising;
 (i′) a sequence of linked nucleosides selected from the group consisting of the native 3′ UTR of any of the nucleic acids that encode any of SEQ ID NOs 8922-17687, SEQ ID NOs 5-21 and functional variants thereof; and 
 (ii′) a 3′ tailing sequence of linked nucleosides. 
   
     
     
         2 . The isolated polynucleotide of  claim 1  wherein the first region of linked nucleosides comprises at least an open reading frame of a nucleic acid sequence, wherein the nucleic acid sequence selected from the group consisting of SEQ ID NOs: 17688-61633. 
     
     
         3 . The isolated polynucleotide of  claim 1 , wherein the 3′ tailing sequence of linked nucleosides is selected from the group consisting of a poly-A tail of approximately 160 nucleotides and a polyA-G quartet. 
     
     
         4 . The isolated polynucleotide of  claim 1  which is purified. 
     
     
         5 . The isolated polynucleotide of  claim 1 , wherein the at least one 5′ terminal cap is selected from the group consisting of Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine. 
     
     
         6 . The isolated polynucleotide of  claim 1 , wherein at least one of said linked nucleosides comprises at least one modification as compared to the chemical structure of an A, G, U or C ribonucleotide. 
     
     
         7 . The isolated polynucleotide of  claim 6 , wherein at least one said modification is located in a nucleoside base and/or sugar portion. 
     
     
         8 - 37 . (canceled) 
     
     
         38 . A pharmaceutical composition comprising the isolated polynucleotide of  claim 1  and a pharmaceutically acceptable excipient. 
     
     
         39 - 40 . (canceled) 
     
     
         41 . A method of producing a polypeptide of interest in a mammalian cell, tissue or organism comprising administering to said cell, tissue or organism the pharmaceutical composition of  claim 38 . 
     
     
         42 . The method of  claim 41 , wherein the isolated polynucleotide is formulated. 
     
     
         43 . The method of  claim 42 , wherein the formulation comprises a lipid. 
     
     
         44 - 50 . (canceled) 
     
     
         51 . A method for producing an increased level of a polypeptide of interest selected from the group consisting of SEQ ID NOs 8922-17687 in a mammalian cell, tissue or organism, comprising administering to said cell, tissue or organism a total daily dose of the pharmaceutical composition of  claim 38  in two or more equal or unequal split doses. 
     
     
         52 - 76 . (canceled)