Kingsman, Alan John; Kingsman, Susan Mary;

Therapeutic Gene

Polynucleotide sequences and vectors containing them, for use in gene therapy, the polynucleotide sequences comprising two or more therapeutic genes operably linked to a promoter and encoding a fusion protein product of the therapeutic genes. The fusion protein may be for example a tyrosine hydroxylase (TH)-DOPA decarboxylase (DD) fusion in either TH-DD or DD-TH order, useful for treating Parkinson's disease.

• Bottle and nipple brush Electromagnetic fuel injection valve
• Solid-state imaging apparatus Lubricating grease
• Modular floating structure Voltage dividing resistor device
• Locking door handle Antibiotic TAN-420
• Orthodontic appliance Magnetic tape cassette
• Wideband short slot hybrid coupler Electric component feeder
• Infrared intrusion detector Operating table
• Double bevel gear angular drive Dust protector
• High-speed cycle clock-synchrounous memory device USB legacy support system
• Grain carrying vehicle Hydrostatic transmission control system
• Concrete reinforcement device Semiconductor pressure sensor
• Pilot valve Rotary body position control apparatus
• Seismometer 2,2-Difluoro-trans-4,5,13,14-tetrahydro-PGI.sub.1 compounds
• Magnetic field generating assembly Firearm
• Three dimensional modelling apparatus Yarn tension control system
• Bypass and unloader valve Ultrasonic treatment of animals
• Slidable door structure Electromagnetic induction air heater
• Image blur preventing device Air action game
• Fiber optic moisture analysis probe Linear deposition apparatus
• Piezoelectric speaker Tuning fork flexural quartz resonator
• Drill tool for deep wells Disposable double vial syringe
• Methods for chromosome-specific staining Centering balloon catheter
• Balance shaft assembly Image heating apparatus
• Megaphone apparatus Folding chair
• Original image projecting apparatus Rotor for an electric motor
• Trailer hitch Adjustable stepladder
• Z-bracket camper jack mounting assembly Solvent reclamation apparatus
• Electrical plug connection Wheeled tool carrier
• Apparatus for producing coils Polymeric compounds
• Flange for a duct

What is claimed is:

1. A lentiviral-based vector capable of transducing a non-dividing cell, said vector comprising a sequence encoding a fusion protein, said sequence comprising two or more therapeutic genes operably linked to a promoter.

2. The vector according to claim 1, wherein the fusion protein is encoded by a sequence comprising tyrosine hydroxylase and a sequence encoding DOPA decarboxylase, wherein the sequences encoding tyrosine hydroxylase and DOPA decarboxylase are operably linked by a sequence encoding a flexible linker.

3. The vector according to claim 1, wherein the lentiviral-based vector comprises a lentviral LTR-deleted vector.

4. The polynucleotide sequence according to claim 1, wherein the flexible linker comprises glycine-serine amino acid repeats.

5. The vector according to claim 1, comprising a single transcription unit comprising the sequences encoding tyrosine hydroxylase and DOPA decarboxylase.

6. A method for producing a bifunctional fusion protein in a non-dividing cell, said method comprising:

providing a non-dividing cell;

transducing the non-dividing cell with the vector according to claim 1; and

expressing the bifunctional fusion protein in the cell, wherein the bifunctional fusion protein comprises tyrosine hydroxylase and DOPA decarboxylase.

7. A target cell in vitro, comprising the lentiviral vector of claim 1.

8. The vector according to claim 5, wherein the single transcription unit is under transcriptional control of the 5' LTR.

9. A lentiviral vector comprising a sequence encoding a fusion protein comprising tyrosine hydroxylase and DOPA decarboxylase, wherein tyrosine hydroxylase and DOPA decarboxylase are operably connected by a flexible linker.

10. A method for producing a bifunctional fusion protein comprising tyrosine hydroxylase and DOPA decarboxylase in a neuron, said method comprising:

providing a neuron;

transducing the neuron with the vector according to claim 9; and

expressing the bifunctional fusion protein in the neuron, wherein the bifunctional fusion protein possesses the activity of both tyrosine hydroxylase and DOPA decarboxylase.

11. The method according to claim 6, wherein the non-dividing cell is a neuron.