Human tyrosine hydroxylase promoter and uses thereof

A method of selecting TH positive cells comprising the steps of:

• a) providing a construct comprising a hTH promoter nucleic acid sequence operably linked to a heterologous nucleic acid sequence;
• b) transfecting a plurality of cells with the construct;
• and
• c) selecting cells expressing the heterologous nucleic acid sequence.

A method of selecting TH positive cells comprising the steps of:

• a) providing a construct comprising an hTH promoter nucleic acid sequence operably linked to a heterologous nucleic acid;
• b) transfecting a plurality of cells with the construct;
• c) differentiating the transfected cells in culture;
• and
• d) selecting cells expressing the heterologous nucleic acid sequence.

A method of treating an individual in need thereof comprising:

• a) providing a construct comprising an hTH promoter nucleic acid sequence operably linked to a heterologous nucleic acid encoding a therapeutic molecule;
• b) transfecting a plurality of cells with the construct;
• c) differentiating the transfected cells in culture;
• d) selecting the differentiated cells expressing the nucleic acid encoding the therapeutic molecule;
• and
• e) administering the selected cells to the individual.

A method of treating an individual in need thereof comprising:

• a) providing a construct comprising an hTH promoter nucleic acid sequence or a functional fragment thereof operably linked to a heterologous nucleic acid encoding a reporter molecule;
• b) transfecting a plurality of cells with the construct;
• c) selecting the cells expressing the nucleic acid encoding the reporter molecule;
• and
• d) administering the selected cells to the individual.

A method of screening for agents affecting the function of a hTH promoter or a functional fragment thereof comprising the steps of:

• a) providing a cell line containing a hTH promoter nucleic acid sequence operably linked to a heterologous nucleic acid encoding a reporter molecule;
• b) contacting said cell line with a candidate agent;
• and
• c) detecting expression of the reporter molecule, wherein increase in the amount of expression of the reporter molecule is indicative of an agent capable of increasing the activity of hTH promoter or a functional fragment thereof and decrease in the amount of expression of the reporter molecule is indicative of an agent capable of decreasing the activity of hTH promoter or a functional fragment thereof.

An isolated nucleic acid sequence comprising nucleic acid sequence or a combination of nucleic acid sequences selected from the group consisting of

• nucleic acids from −10,761 to −10,752 of the +strand of the hTH promoter of SEQ ID NO:
• 1;
• nucleic acids from −8,605 to −8,596 of the +strand of the hTH promoter of SEQ ID NO:
• 1;
• nucleic acids from −8,180 to −8,171 of the +strand of the hTH promoter of SEQ ID NO:
• 1;
• nucleic acids from −4,075 to −4,084 of the −strand of the hTH promoter of SEQ ID NO:
• 1;
• nucleic acids from −2,584 to −2,575 of the +strand of the hTH promoter of SEQ ID NO:
• 1;
• nucleic acids from −8,917 to −8,876 of the +strand of the hTH promoter of SEQ ID NO:
• 1;
• nucleic acids from −7,248 to −7,207 of the +strand of the hTH promoter of SEQ ID NO:
• 1;
• nucleic acids from −5,489 to −5,472 of the +strand of the hTH promoter of SEQ ID NO:
• 1;
• nucleic acids from −5,399 to −5,349 of the +strand of the hTH promoter of SEQ ID NO:
• 1;
• and
• nucleic acids from −2,423 to −2,327 of the +strand of the hTH promoter of SEQ ID NO:
• 1
• wherein the nucleic acid is operably linked to a heterologous nucleic acid sequence.