title: Establishment and characterization of CAG/EGFP transgenic rabbit line.
abstract: Cell marking is a very important procedure for identifying donor cells after cell and/or organ transplantation in vivo. Transgenic animals expressing marker proteins such as enhanced green fluorescent protein (EGFP) in their tissues are a powerful tool for research in fields of tissue engineering and regenerative medicine. The purpose of this study was to establish transgenic rabbit lines that ubiquitously express EGFP under the control of the cytomegalovirus immediate early enhancer/beta-actin promoter (CAG) to provide a fluorescent transgenic animal as a bioresource. We microinjected the EGFP expression vector into 945 rabbit eggs and 4 independent transgenic candidate pups were obtained. Two of them died before sexual maturation and one was infertile. One transgenic male candidate founder rabbit was obtained and could be bred by artificial insemination. The rabbit transmitted the transgene in a Mendelian manner. Using fluorescence in situ hybridization analysis, we detected the transgene at 7q11 on chromosome 7 as a large centromeric region in two F1 offspring (one female and one male). Eventually, one transgenic line was established. Ubiquitous EGFP fluorescence was confirmed in all examined organs. There were no gender-related differences in fluorescence. The established CAG/EGFP transgenic rabbit will be an important bioresource and a useful tool for various studies in tissue engineering and regenerative medicine.
authors.affiliation: The YS Institute Inc., 1198-4 Iwazo, Utsunomiya, Tochigi, Japan. ystg@ystg.co.jp
authors.forename: Ri-ichi
authors.initials: R
authors.lastname: Takahashi
authors.forename: Takashi
authors.initials: T
authors.lastname: Kuramochi
authors.forename: Kazuki
authors.initials: K
authors.lastname: Aoyagi
authors.forename: Shu
authors.initials: S
authors.lastname: Hashimoto
authors.forename: Ichiro
authors.initials: I
authors.lastname: Miyoshi
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kasai
authors.forename: Yoji
authors.initials: Y
authors.lastname: Hakamata
authors.forename: Eiji
authors.initials: E
authors.lastname: Kobayashi
authors.forename: Masatsugu
authors.initials: M
authors.lastname: Ueda
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001947:Breeding; D005260:Female; D005453:Fluorescence; D015870:Gene Expression; D005818:Genetic Engineering; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D011817:Rabbits / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins