Genetic element that enhances protein translation

• US 9,234,197 B2
• Filed: 10/26/2012
• Issued: 01/12/2016
• Est. Priority Date: 10/28/2011
• Status: Active Grant

• Current Assignee: Arizona Board of Regents (University of Arizona)
• Original Assignee: Arizona Board of Regents (University of Arizona)
• Inventors: Chaput, John, Jacobs, Bertram, Wellensiek, Brian, Flores, Julia
• Primary Examiner(s): Leith, Nancy J
• Application Number:
  US14/349,835
• Publication Number:
  US 20140255990A1
• Time in Patent Office: 1,173 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2830/30 being an enhancer not forming part of the promoter region
• CPC Class Codes: C12N 2830/85 mammalian
• CPC Class Codes: C12N 2840/105 enhancing translation
• CPC Class Codes: C12N 2840/85 mammalian

Abstract

The present invention provides isolated polynucleotides that can serve as translation enhancing elements and their use in protein expression reagents and methods.

Claims

1. A recombinant viral or plasmid expression vector comprising

(a) a translation enhancement element (TEE) of between 37 and 42 nucleotides in length, comprising the nucleic acid sequence of 5′

-CATATTGAAGAGACAGAGTGATATATAAAACTGCTAA-3′

(SEQ ID NO;

     1); and

(b) a cloning site located downstream of the TEE.

2. The expression vector of claim 1, further comprising a promoter located upstream of the TEE.

3. The expression vector of claim 1, further comprising a protein encoding nucleic acid cloned into the cloning site.

4. An isolated recombinant host cell comprising the expression vector of claim 1.

5. A method for protein expression, comprising contacting the expression vector of claim 3 with reagents and under conditions for promoting expression of the protein encoded by the protein-encoding nucleic acid, and expressing the protein.

6. The method of claim 5, wherein the protein is expressed in a cell-free translation system.

7. The method of claim 5, wherein the protein is expressed by a recombinant host cell.

8. The method of claim 5, wherein the expression vector is a recombinant vaccinia virus.

9. The method of claim 7, wherein the recombinant host cell expresses the protein using non-viral-mediated translation.

10. An isolated recombinant virus, comprising the expression vector of claim 1, wherein the expression vector is comprises a viral expression vector.

11. The recombinant virus of claim 10, wherein the recombinant virus is a vaccinia virus, and the viral expression vector is a vaccinia virus expression vector.

12. The expression vector of claim 1, wherein the TEE consists of 5′

-CATATTGAAGAGACAGAGTGATATATAAAACTGCTAA-3′

(SEQ ID NO;

     1).

13. The expression vector of claim 1, wherein the TEE consists of 5′

-AGAACCATATTGAAGAGACAGAGTGATATATAAAACTGCTAA-3′

(SEQ ID NO;

2).