US 10,472,638 B2Granted

Synthetic MRNA leaders

AssigneeVectron Biosolutions AS

InventorsRahmi Lale, Svein Valla, Simone Balzer Le

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Abstract

The present invention provides a synthetic mRNA leader for enhancing the expression of a gene encoding a desired protein, vectors comprising said synthetic mRNA leader and methods of producing a desired gene product using said synthetic mRNA leader and vector, said method comprising expressing said gene using a synthetic mRNA leader which comprises from 5′ to 3′: (i) a first mRNA leader sequence element; (ii) a spacer region; and (iii) a second mRNA leader sequence element; wherein said first mRNA leader sequence element is a modified transcription-stimulating mRNA leader capable of enhancing transcription of a gene relative to an unmodified reference mRNA leader sequence and/or said second mRNA leader element is a modified translation-stimulating mRNA leader capable of enhancing the translation of a gene transcript relative to an unmodified reference mRNA leader sequence.

Claims16

Dependent claims

A method of enhancing expression of a desired gene product in a recombinant gene expression system, said method comprising expressing said gene using a synthetic mRNA leader which comprises from 5′ to 3′:

(i) a first mRNA leader sequence element;
(ii) a spacer region;
and
(iii) a second mRNA leader sequence element;
wherein said first mRNA leader sequence element is a first mutated Pm mRNA leader capable of enhancing transcription of a gene relative to an unmutated Pm mRNA leader sequence of SEQ ID NO:1, wherein the first mutated Pm mRNA leader has at least 80% sequence identity to the unmutated Pm mRNA leader sequence of SEQ ID NO:1;
and
wherein said second mRNA leader sequence element is a second mutated Pm mRNA leader capable of enhancing the translation of a gene transcript relative to an unmutated Pm mRNA leader sequence of SEQ ID NO:1, wherein the second mutated Pm mRNA leader has at least 80% sequence identity to the unmutated Pm mRNA leader sequence of SEQ ID NO:1.

4 more elements

Dependencies are inferred from the claim text.

Facts

Application: 15/300,993Filed 2 Apr 2015
Priority: GB 1406006.53 Apr 2014
Published: US 2017/0022509 A126 Jan 2017
Granted: 12 Nov 2019
Term ends: Not estimatedThe source records don’t support an estimate.

Classification7 codes

C12N 15/67
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression: general methods for enhancing…
C12N 15/63
Recombinant DNA-technology: introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
C12N 15/70
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression: vectors or expression systems…
C12N 15/78
Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora: for Pseudomonas