Development of a unique system for spatiotemporal and lineage-specific gene expression in mice.
We have developed an advanced method for conditional gene expression in mice that integrates the Cre-mediated and tetracycline-dependent expression systems. An rtTA gene, preceded by a loxP-flanked STOP sequence, was inserted into the ROSA26 locus to create a R26STOPrtTA mouse strain. When the STOP sequence is excised by Cre-mediated recombination, the rtTA is expressed in the Cre-expressing cells and all of their derivatives. Therefore, cell type-, tissue-, or lineage-specific expression of rtTA is achieved by the use of an appropriate Cre transgenic strain. In mice also carrying a target gene under the control of the tetracycline response element, inducible expression of the target gene is temporally regulated by administration of doxycycline. Our results demonstrate that this universal system is uniquely suited for spatiotemporal and lineage-specific gene expression in an inducible fashion. Gene expression can be manipulated in specific cell types and lineages with a flexibility that is difficult to achieve with conventional methods.
Yu
Hsiao-Man Ivy
HM
Center for Oral Biology, Department of Biomedical Genetics, Abs Institute of Biomedical Sciences, School of Medicine and Dentistry, University of Rochester, 601 Elmwood Avenue, Box 611, Rochester, NY 14642, USA.
Liu
Bo
B
Chiu
Shang-Yi
SY
Costantini
Frank
F
Hsu
Wei
W
D000818:Animals; D004318:Doxycycline / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins; D019426:Integrases / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009432:Neural Crest / Q000378:metabolism; D011506:Proteins / Q000235:genetics*; D022661:RNA, Untranslated; D013752:Tetracycline / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D001616:beta-Galactosidase
C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D011506:Proteins; D022661:RNA, Untranslated; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases; D001616:beta-Galactosidase; D013752:Tetracycline; D004318:Doxycycline