Long-lived polynucleotide molecules

Abstract

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

1 . An isolated codon optimized mRNA, wherein said mRNA comprising a nucleic acid sequence encoding
 (a) a polypeptide of interest; and   (b) a first longevity enhancing sequence.   
     
     
         2 . The isolated codon optimized mRNA of  claim 1  wherein the first longevity enhancing sequence is selected from the group consisting of carboxy-terminal peptide, albumin and IgG4. 
     
     
         3 . The isolated codon optimized mRNA of  claim 1  wherein at least one isolated mRNA comprises at least a first modified nucleoside. 
     
     
         4 . The isolated codon optimized mRNA of  claim 1  comprising a poly-A tail and wherein the poly-A tail is at least 140 nucleotides in length. 
     
     
         5 . The isolated codon optimized mRNA of  claim 1  comprising a 5′ cap structure and wherein the 5′ cap structure 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 codon optimized mRNA of  claim 1 , wherein the first longevity enhancing sequence is located at a position in the mRNA selected from the group consisting of 5′UTR, coding region and 3′UTR. 
     
     
         7 . The isolated codon optimized mRNA of  claim 1 , wherein the first longevity enhancing sequence is a carboxy-terminal peptide and is selected from the group consisting of SEQ ID NO: 1-3. 
     
     
         8 . The isolated codon optimized mRNA of  claim 7  encoding a second longevity enhancing sequence. 
     
     
         9 . The isolated codon optimized mRNA of  claim 8 , wherein the second longevity enhancing sequence is a carboxy-terminal peptide and is selected from the group consisting of SEQ ID NO: 1-3. 
     
     
         10 . The isolated codon optimized mRNA of  claim 8 , wherein the first longevity enhancing sequence and the second longevity enhancing sequence are the same. 
     
     
         11 . The isolated codon optimized mRNA of  claim 8  encoding a third longevity enhancing sequence. 
     
     
         12 . The isolated codon optimized mRNA of  claim 11 , wherein the third longevity enhancing sequence is a carboxy-terminal peptide and is selected from the group consisting of SEQ ID NO: 1-3. 
     
     
         13 . The isolated codon optimized mRNA of  claim 11 , wherein the first longevity enhancing sequence, the second longevity enhancing sequence and the third longevity enhancing sequence are the same. 
     
     
         14 . The isolated codon optimized mRNA of  claim 11 , wherein the first longevity enhancing sequence is located in the 5′UTR of said isolated codon optimized mRNA. 
     
     
         15 . The isolated codon optimized mRNA of  claim 14 , wherein the second longevity enhancing sequence and the third longevity enhancing sequence are located in the 3′UTR of said isolated codon optimized mRNA. 
     
     
         16 . A method of increasing the half-life of a protein in a cell, tissue and/or organism comprising contacting the cell, tissue and/or organism with the isolated codon optimized mRNA of  claim 1 . 
     
     
         17 . A pharmaceutical composition comprising the isolated codon optimized mRNA of  claim 1  and a pharmaceutically acceptable excipient.