Construction and characterization of a doxycycline-inducible transgenic system in Msx2 expressing cells.
Homeobox gene Msx2 is widely expressed during both embryogenesis and postnatal development and plays important roles during organogenesis. We developed an Msx2-rtTA BAC transgenic line which can activate TetO-Cre expression in Msx2-expressing cells upon doxycycline (Dox) treatment. Using the Rosa26-LacZ (R26R) reporter line, we show that rtTA is activated in Msx2-expressing organs including the limb, heart, external genitalia, urogenital system, hair follicles and craniofacial regions. Moreover, we show that in body appendages, the transgene can be activated in different domains depending on the timing of Dox treatment. In addition, the transgene can also be effectively activated in adult tissues such as the hair follicle and the urogenital system. Taken together, this Msx2-rtTA;TetO-Cre system is a valuable tool for studying gene function in the development of the aforementioned organs in a temporal and spatially-restricted manner, as well as for tissue lineage tracing of Msx2-expressing cells. When induced postnatally, this system can also be used to study gene function in adult tissues without compromising normal development and patterning.
Lin
Congxing
C
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri, USA.
Yin
Yan
Y
Chen
Hong
H
Fisher
Alexander V
AV
Chen
Feng
F
Rauchman
Michael
M
Ma
Liang
L
D000818:Animals; D004318:Doxycycline / Q000494:pharmacology*; D004622:Embryo, Mammalian / Q000196:embryology / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D010078:Oxazines; D010957:Plasmids / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D022661:RNA, Untranslated; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors; D019076:Transgenes / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D018398:Homeodomain Proteins; C084119:MSX2 protein; D010078:Oxazines; D011506:Proteins; D022661:RNA, Untranslated; C008619:Nile Blue; D001616:beta-Galactosidase; D004318:Doxycycline