Local Cre-mediated gene recombination in vascular smooth muscle cells in mice.
Here we describe a means to conditionally modify genes at a predefined and localized region of the vasculature using a perivascular drug delivery device (PDD). A 4-hydroxytamoxifen (4-OHT)-eluting PDD was applied around the carotid or femoral artery of a mouse strain carrying both the tamoxifen-inducible and smooth muscle cell (SMC)-specific Cre-recombinase (SM-Cre-ER(T2)) transgene and a stop-floxed beta-galactosidase gene in the Rosa26 locus: the SM-CreER(T2)(ki)/rosa26 mouse. A dose and time curve of 0-10% (w/w) 4-OHT and 0-14 days application of the PDD in SM-CreER(T2)(ki)/rosa26 mice showed optimal gene recombination at 1% (w/w) 4-OHT loading at 7 days post application (carotid artery 2.4+/-1.8%; femoral artery 4.0+/-3.8% of SMCs). The unique 4-OHT-eluting PDD allowed us to achieve SMC-specific recombination in the same order of magnitude as compared to systemic tamoxifen administration. In addition, recombination was completely confined to the PDD-treated vessel wall segment. Thus, local application of a 4-OHT-eluting PDD results in vascular SMC-specific Cre-mediated recombination in SM-CreER(T2)(ki)/rosa26 mice without affecting additional SMCs.
Department of Cardiology, p/o TNO-Quality of Life Gaubius Laboratory, Leiden University Medical Centre, Zernikedreef 9, P.O. Box 2215, 2301 CE, Leiden, The Netherlands. asm.zadelaar@pg.tno.nl
Susanne M
SM
Zadelaar
Lianne S M
LS
Boesten
Nuno M M
NM
Pires
Anita
A
van Nieuwkoop
Erik A L
EA
Biessen
Wouter
W
Jukema
Louis M
LM
Havekes
Bart J M
BJ
van Vlijmen
Ko
K
Willems van Dijk
D000818:Animals; D016503:Drug Delivery Systems; D019426:Integrases / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic / Q000187:drug effects; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology
D013629:Tamoxifen; C016601:afimoxifene; C045073:Cre recombinase; D019426:Integrases