US2004086980A1PendingUtilityA1

Methods for producing nucleic acids lacking 3'-untranslated regions and optimizing cellular RNA-protein fusion formation

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Priority: Aug 17, 1998Filed: Aug 21, 2003Published: May 6, 2004
Est. expiryAug 17, 2018(expired)· nominal 20-yr term from priority
Inventors:Philip W. HammondDasa Lipovsek
C12N 15/10C12N 15/1062C12N 15/1075C12N 15/1096
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Claims

Abstract

Described herein are methods for removing the 3′-untranslated regions from cDNA or mRNA molecules, as well as methods for the use of such products for RNA-protein fusion formation.

Claims

exact text as granted — not AI-modified
1 . A method for removing the 3′-untranslated region of a population of DNA molecules, wherein each DNA molecule in said population of DNA molecules comprises an open reading frame and a 3′-untranslated region, said method comprising: 
 (a) providing a population of DNA molecules, each of said DNA molecules terminating at its 5′ end in an overhang and at its 3′ end in a blunt end; and  
 (b) treating each of said DNA molecules first with a 3′→5′ exonuclease and then with a single-stranded nuclease under conditions that allow removal of said 3′-untranslated region.  
 
     
     
         2 . The method of  claim 1 , wherein said 3→5′ exonuclease is exonuclease III.  
     
     
         3 . The method of  claim 1 , wherein said nuclease is Mung bean nuclease.  
     
     
         4 . The method of  claim 1 , wherein step (b) further results in removal of the stop codon of said open reading frame.  
     
     
         5 . The method of  claim 1 , wherein each of said DNA molecules is a cDNA produced by reverse transcription from an mRNA sequence.  
     
     
         6 . The method of  claim 1 , wherein said population comprises at least 10 DNA molecules.  
     
     
         7 . The method of  claim 1 , wherein said population comprises at least 10 2  DNA molecules.  
     
     
         8 . The method of  claim 1 , wherein said population comprises at least 10 3  DNA molecules.  
     
     
         9 . The method of  claim 1 , wherein said population comprises at least 10 4  DNA molecules.  
     
     
         10 . The method of  claim 1 , wherein said population comprises at least 10 5  DNA molecules.  
     
     
         11 . The method of  claim 1 , wherein said population comprises at least 10 6  DNA molecules.

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