L-DOPA AND DOPAMINE-PRODUCING GENE CASSETTES FOR GENE-THERAPY APPROACHES TO PARKINSONS-DISEASE

As an aid in the development of vector systems for use in gene therapy paradigms of central nervous system disorders such as Parkinson's dis ease, we have developed L-Dopa or dopamine-producing gene cassettes. S pecifically, a human tyrosine hydroxylase cDNA (HTH-S) was rendered co nstitutively active by truncation of the N-terminal regulatory domain (tHTH). In addition, a bicistronic construct capable of directing the production of dopamine was created by inserting an internal ribosome e ntry site downstream of tHTH followed by the coding sequences of aroma tic amino acid decarboxylase. All three constructs generated immunorea ctive peptides of the predicted size, were enzymatically active, and p roduced L-Dopa (HTH-2, tHTH) or dopamine (bicistronic construct) follo wing transient transfection of COS-7 cells. These constructs, in conju nction with viral or nonviral expression systems, may be efficacious i n gene therapy approaches to Parkinson's disease. (C) 1997 Academic Pr ess.