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(Circulation Research. 2001;88:587.)
© 2001 American Heart Association, Inc.

Integrative Physiology

Inducible Gene Targeting in Postnatal Myocardium by Cardiac-Specific Expression of a Hormone-Activated Cre Fusion Protein

Tetsuo Minamino, Vinciane Gaussin, Francesco J. DeMayo, Michael D. Schneider

From the Center for Cardiovascular Development (T.M., V.G., M.D.S.) and Departments of Medicine (T.M., V.G., F.J.D., M.D.S.), Molecular and Cellular Biology (M.D.S.), and Molecular Physiology and Biophysics (M.D.S.), Baylor College of Medicine, Houston, Tex. Present address of V.G. is Cardiovascular Research Institute, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, and Hackensack University Medical Center, Hackensack, NJ.

Correspondence to Dr Michael D. Schneider, Center for Cardiovascular Development, Baylor College of Medicine, One Baylor Plaza, Room 506C, Houston, TX 77030. E-mail michaels{at}bcm.tmc.edu

Abstract—Cardiac-restricted expression of Cre recombinase can provoke lineage-specific gene excision in the myocardium. However, confounding early lethality may still preclude using loss-of-function models to study the postnatal heart. Here, we have tested whether inducible, heart-specific recombination can be triggered after birth by transgenic expression of a Cre fusion protein that incorporates a mutated progesterone receptor ligand binding domain (PR1) that is activated by the synthetic antiprogestin, RU486, but not by endogenous steroid hormones. CrePR1 driven by the -myosin heavy chain (MHC) promoter was expressed specifically in heart. Translocation of CrePR1 from cytoplasm to nuclei in ventricular myocytes was induced by RU486. To establish whether this approach can mediate cardiac-specific, drug-dependent excision between loxP sites in vivo, we mated MHC-CrePR1 mice with a ubiquitously expressed (ROSA26) Cre reporter line. Offspring harboring MHC-CrePR1 and/or the floxed allele were injected with RU486 versus vehicle, and the prevalence of ß-galactosidase (ß-gal)–positive cells was determined, indicative of Cre-mediated excision. Little or no baseline recombination was seen 1 week after birth. Cardiac-restricted, RU486-inducible recombination was demonstrated in bigenic mice at age 3 and 6 weeks, using each of 3 independent CrePR1 lines. Recombination in the absence of ligand paralleled the levels of CrePR1 protein expression and was more evident at 6 weeks. Thus, conditional, posttranslational activation of a Cre fusion protein can bypass potential embryonic and perinatal effects on the heart and permits inducible recombination in cardiac muscle. High levels of the chimeric Cre protein, in particular, were associated with progressive recombination in the absence of drug.

Key Words: Cre recombinase • genetics • progesterone receptor • transgenic mice

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