title: Generation of Ci-Brachyury-GFP stable transgenic lines in the ascidian Ciona savignyi.
abstract: We report generation of stable transgenic lines of the ascidian Ciona savignyi carrying a Ciona intestinalis-Brachyury-promoter/Green Fluorescent Protein-reporter (Ci-Bra-GFP) construct. The transgenic lines were made using a technique in which the endonuclease I-SceI was coinjected into fertilized eggs with a transgene construct containing flanking recognition sites for I-SceI. Two founder animals, out of 12 F(0) adults tested, were found to transmit the transgene to their offspring (F(1)s) at frequencies of 42% and 23%. The transgene was further inherited by the F(2) in a Mendelian fashion and displayed nonmosaic expression, indicating integration into the genome. The Mendelian inheritance and the absence of mosaicism persisted through the F(3) and F(4) generations. Southern blot analyses showed that the transgene was organized in tandem arrays of no more than 10 copies. Using these Ci-Bra-GFP transgenics, we describe cellular movements and shape changes involved in notochord morphogenesis in both wildtype and mutant embryos.
authors.affiliation: Molecular Cellular and Developmental Biology Department, Neuroscience Research Institute, University of California, Santa Barbara, CA 93106, USA.
authors.forename: Karine
authors.initials: K
authors.lastname: Deschet
authors.forename: Yuki
authors.initials: Y
authors.lastname: Nakatani
authors.forename: William C
authors.initials: WC
authors.lastname: Smith
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D015139:Blotting, Southern; D005326:Fetal Proteins / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D009030:Mosaicism; D009154:Mutation; D009672:Notochord / Q000196:embryology / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020825:T-Box Domain Proteins / Q000235:genetics* / Q000378:metabolism; D019076:Transgenes; D014561:Urochordata / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D000098908:Brachyury Protein
chemical_list: D005326:Fetal Proteins; D020825:T-Box Domain Proteins; D000098908:Brachyury Protein