Chimeric post-transcriptional regulatory element

Abstract

The present disclosure relates to chimeric post-transcriptional regulatory elements (PRE) and vectors useful for expressing a protein in a cell. The PRE contains alpha, beta and optionally gamma subelements selected from different native PRE sequences and are discovered to be more potent than their native counterparts.

1 - 15 . (canceled) 
     
     
         16 . A polynucleotide comprising:
 (a) a first fragment consisting of the nucleic acid sequence of SEQ ID NO: 5, 9, or 18, or a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 5, 9 or 18, and   (b) a second fragment consisting of the nucleic acid sequence of SEQ ID NO: 3 or a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 3.   
     
     
         17 . The polynucleotide of  claim 16 , further comprising (c) a third fragment consisting of a gamma subelement of a post-transcriptional regulatory element (PRE). 
     
     
         18 - 20 . (canceled) 
     
     
         21 . A polynucleotide construct, comprising (a) a first fragment consisting of the nucleic acid sequence of SEQ ID NO: 5, 9, or 18, or a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 5, 9 or 18, (b) a second fragment consisting of the nucleic acid sequence of SEQ ID NO: 3 or a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 3 and (c) a protein-coding sequence. 
     
     
         22 . The polynucleotide construct of  claim 21 , wherein the protein-coding sequence is located between the first fragment and the second fragment. 
     
     
         23 . The polynucleotide construct of  claim 21 , further comprising a 3′-UTR. 
     
     
         24 . The polynucleotide construct of  claim 23 , wherein the 3′-UTR is located between the first fragment and the second fragment. 
     
     
         25 . The polynucleotide construct of  claim 21 , further comprising a poly(A) sequence. 
     
     
         26 . An isolated cell comprising the polynucleotide construct of  claim 21 . 
     
     
         27 . The polynucleotide of claim  18 , wherein the first fragment consists of the nucleic acid sequence of SEQ ID NO: 9, or a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 9. 
     
     
         28 . The polynucleotide construct of  claim 21 , wherein the first fragment consists of the nucleic acid sequence of SEQ ID NO: 9, or a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 9.