Published online before print March 6, 2003, doi: 10.1161/01.RES.0000065442.64694.9F
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© 2003 American Heart Association, Inc.
Molecular Medicine |
Reengineering Inducible Cardiac-Specific Transgenesis With an Attenuated Myosin Heavy Chain Promoter
From the Department of Pediatrics, Division of Molecular Cardiovascular Biology (A.S., J.G., Q.L., H.O., J.R.), The Children’s Hospital Research Foundation, Cincinnati, Ohio, and Procter and Gamble Pharmaceuticals (M.C.H.), Health Care Research Center-Discovery, Mason, Ohio.
Correspondence to Jeffrey Robbins, Department of Pediatrics, Division of Molecular Cardiovascular Biology, MLC7020 The Children’s Hospital Research Foundation, Cincinnati, OH 45229-3039. E-mail jeff.robbins{at}chmcc.org
Despite the advantages of reversibly altering cardiac transgene expression, the number of successful studies with inducible cardiac-specific transgene expression remains limited. The utility of the current system is hampered by the large number of lines needed before a nonleaky inducible line is isolated and by the use of a heterologous virus-based minimal promoter in the responder line. We developed an efficient, experimentally flexible system that enables us to reversibly affect both abundant and nonabundant cardiomyocyte proteins. The use of bacterial-codon–based transactivators led to aberrant splicing, whereas other more efficient transactivators, by themselves, caused disease when expressed in the heart. The redesign of the system focused on developing stable transactivator-expressing lines in which expression was driven by the mouse 
-myosin heavy chain promoter. A minimal responder locus was derived from the same promoter, in which the GATA sites and thyroid responsive elements responsible for robust cardiac specific expression were ablated, leading to an attenuated promoter that could be inducibly controlled. In all cases, whether activated or not, expression mimicked that of the parental promoter. By use of this system, an inducible expression of an abundant contractile protein, the atrial isoform of essential myosin light chain 1, and a powerful biological effector, glycogen synthase kinase-3ß (GSK-3ß), were obtained. Subsequently, we tested the hypothesis that GSK-3ß expression could reverse a preexisting hypertrophy. Inducible expression of GSK-3ß could both attenuate a hypertrophic response and partially reverse a pressure-overload–induced hypertrophy. The system appears to be robust and can be used to temporally control high levels of cardiac-specific transgene expression.
Key Words: transgene • myosin • cardiac disease • muscle • mice
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