Noval modified MSP-1 nucleic acid sequences and methods for increasing mRNA levels and protein expression in cell systems

• US 20050235371A1
• Filed: 05/27/2005
• Published: 10/20/2005
• Est. Priority Date: 10/20/1997
• Status: Active Grant

• Current Assignee: GTC Biotherapeutics, Inc.
• Original Assignee: Genzyme Transgenics Corporation
• Inventors: Meade, Harry M., Chen, Li How
• Granted Patent: US 7,354,594 B2
• Time in Patent Office: Days
• US Class Current: 800/14
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/10 Mammal
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A61K 2039/51 comprising whole cells, viruses or DNA/RNA
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/445 Plasmodium
• CPC Class Codes: C07K 14/4732 Casein in foodstuffs A23J
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides modified recombinant nucleic acid sequences (preferably DNA) and methods for increasing the mRNA levels and protein expression of malarial surface protein MSP-1 which is known to be difficult to express in cell culture systems, mammalian cell culture systems, or in transgenic animals. The preferred protein candidates for expression using the recombinant techniques of the invention are MSP-1 proteins expressed from DNA coding sequences comprising reduced overall AT content or AT rich regions and/or mRNA instability motifs and/or rare codons relative to the native MSP-1 gene.

Claims

1. A modified MSP-1 nucleic acid sequence, as shown in SEQ. ID. NO 1.

4. A vector comprising a modified MSP-1 nucleic acid according to claim 1 and a goat beta casein promoter and signal sequence.

5. A host cell transformed with the vector according to claim 4.

6. A transgenic non-human mammal comprising a vector according to claim 4.

7. A transgenic non-human mammal comprising a modified MSP-1 nucleic acid according to claim 1.

8. A DNA vaccine comprising a modified nucleic acid according to claim 1.

2. A process for preparing a modified MSP-1 nucleic acid comprising the steps of lowering the overall AT content of the natural gene encoding MSP-1, eliminating all mRNA instability motifs and replacing all rare codons with a preferred codon of the mouse mammary gland.

3. A modified MSP-1 nucleic acid prepared by the process according to claim 2.