Gene recombination in postmitotic cells. Targeted expression of Cre recombinase provokes cardiac-restricted, site-specific rearrangement in adult ventricular muscle in vivo.
Open Access
- 1 July 1997
- journal article
- Published by American Society for Clinical Investigation in JCI Insight
- Vol. 100 (1), 169-179
- https://doi.org/10.1172/jci119509
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.Keywords
This publication has 47 references indexed in Scilit:
- Single-copy transgenic mice with chosen-site integration.Proceedings of the National Academy of Sciences of the United States of America, 1996
- Conditional gene targeting.JCI Insight, 1996
- Targeted oncogene activation by site-specific recombination in transgenic mice.Proceedings of the National Academy of Sciences of the United States of America, 1992
- A sensitive method for the detection of beta-galactosidase in transfected mammalian cells.1991
- Tissue-specific regulation of the alpha-myosin heavy chain gene promoter in transgenic mice.Online Journal of Public Health Informatics, 1991
- FVB/N: an inbred mouse strain preferable for transgenic analyses.Proceedings of the National Academy of Sciences of the United States of America, 1991
- Recombinase-Mediated Gene Activation and Site-Specific Integration in Mammalian CellsScience, 1991
- Developmental regulation of myosin gene expression in mouse cardiac muscle.The Journal of cell biology, 1990
- A simple phase-extraction assay for chloramphenicol acyltransferase activityGene, 1988
- A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye BindingAnalytical Biochemistry, 1976