Construction of Synthetic Muscle Specific Promoters

Somatic gene therapy is expected to provide new and improved therapies for many diseases.We are developing a new class of DNA-based therapeutics, "gene medicines" which are expected toprovide unique clinical benefits for the treatment of a broad range of diseases. Gene medicines utilizetissue-specific DNA expression vectors and novel vector delivery systems for the in vivo delivery oftherapeutic proteins. Clinically effective and safe gene medicines will require the development of reliablemethods to regulate gene expression such that synthesis of the therapeutic protein is not only restrictedto the proper cell-type and tissue but is also at a level that remains within the therapeutic rangeappropriate for the disease being treated. Our previous work at our laboratories and in the founder'slaboratories has led to the development of a muscle-specific vector system (MVS) capable of directingthe synthesis of proteins in muscle in vivo. The control of gene expression in eukaryotic cells iscomplex, involving the interaction of a variety of tissue-specific positive and negative trans-acting proteinfactors with specific regulatory sequences associated with tissue-regulated genes. MVS vectorsincorporate upstream and downstream regulatory sequences derived from the well-characterized chickenskeletal alpha-actin gene to control the expression, mRNA stability and translation of clinically relevantproteins. The purpose of this research is to develop synthetic muscle-specific vectors which will providehigher levels of expression in mammalian muscle cells than natural promoter elements. We will constructa series of synthetic promoter/enhancer fragments based on the sequences of transcriptional controlelements involved in the activation and regulation of genes in mammalian muscle cells.