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Research Article Free access | 10.1172/JCI119509

Gene recombination in postmitotic cells. Targeted expression of Cre recombinase provokes cardiac-restricted, site-specific rearrangement in adult ventricular muscle in vivo.

R Agah, P A Frenkel, B A French, L H Michael, P A Overbeek, and M D Schneider

Molecular Cardiology Unit, Baylor College of Medicine, Houston, Texas 77030, USA.

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Molecular Cardiology Unit, Baylor College of Medicine, Houston, Texas 77030, USA.

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Molecular Cardiology Unit, Baylor College of Medicine, Houston, Texas 77030, USA.

Find articles by French, B. in: JCI | PubMed | Google Scholar

Molecular Cardiology Unit, Baylor College of Medicine, Houston, Texas 77030, USA.

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Molecular Cardiology Unit, Baylor College of Medicine, Houston, Texas 77030, USA.

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Molecular Cardiology Unit, Baylor College of Medicine, Houston, Texas 77030, USA.

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Published July 1, 1997 - More info

Published in Volume 100, Issue 1 on July 1, 1997
J Clin Invest. 1997;100(1):169–179. https://doi.org/10.1172/JCI119509.
© 1997 The American Society for Clinical Investigation
Published July 1, 1997 - Version history
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Abstract

Mouse models of human disease can be generated by homologous recombination for germline loss-of-function mutations. However, embryonic-lethal phenotypes and systemic, indirect dysfunction can confound the use of knock-outs to elucidate adult pathophysiology. Site-specific recombination using Cre recombinase can circumvent these pitfalls, in principle, enabling temporal and spatial control of gene recombination. However, direct evidence is lacking for the feasibility of Cre-mediated recombination in postmitotic cells. Here, we exploited transgenic mouse technology plus adenoviral gene transfer to achieve Cre-mediated recombination in cardiac muscle. In vitro, Cre driven by cardiac-specific alpha-myosin heavy chain (alphaMyHC) sequences elicited recombination selectively at loxP sites in purified cardiac myocytes, but not cardiac fibroblasts. In vivo, this alphaMyHC-Cre transgene elicited recombination in cardiac muscle, but not other organs, as ascertained by PCR analysis and localization of a recombination-dependent reporter protein. Adenoviral delivery of Cre in vivo provoked recombination in postmitotic, adult ventricular myocytes. Recombination between loxP sites was not detected in the absence of Cre. These studies demonstrate the feasibility of using Cre-mediated recombination to regulate gene expression in myocardium, with efficient induction of recombination even in terminally differentiated, postmitotic muscle cells. Moreover, delivery of Cre by viral infection provides a simple strategy to control the timing of recombination in myocardium.

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