SYNTHETIC MUSCLE PROMOTERS WITH ACTIVITIES EXCEEDING NATURALLY OCCURRING REGULATORY SEQUENCES IN CARDIAC CELLS

摘要

Transgenes driven by naturally occurring cardiac promoters have relatively lowlevels of cardiac transgenic gene expression, and have consequently limitedthe use of cardiac muscle as a target for plasmid mediated genesupplementation. However, by randomly assembling motifs of E-box, MEF-2, TEF-1and SRE elements, cardiac-specific synthetic promoter recombinant librarieshave been produced. By screening hundreds of resultant clones fortranscriptional activity both in vitro and in vivo, a few cardiac-specificsynthetic promoters were discovered comprising a transcriptional potency thatgreatly exceeds the transcriptional levels obtained from natural myogenic andviral gene promoters. These promoters are used to direct the expression ofdesirable genes in nucleic acid expression constructs specifically to cardiaccells. Thus, these cardiac specific-synthetic promoters can be utilized forplasmid mediated gene supplementation for serious health conditions, such asischemic disease, myocardial infarction or heart failure. Thus, one aspect ofthe current invention is a cardiac specific-synthetic promoter produced by amethod that generates a library of randomized synthetic-promoter-recombinantexpression constructs. Another aspect of the present invention is directed toa method using the cardiac specific-synthetic expression construct forexpression a gene of interest in a cardiac cell.