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Generation of Ci‐Brachyury‐GFP stable transgenic lines in the ascidian Ciona savignyi
Author(s) -
Deschet Karine,
Nakatani Yuki,
Smith William C.
Publication year - 2003
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.10195
Subject(s) - biology , transgene , brachyury , green fluorescent protein , notochord , ciona intestinalis , ciona , mcherry , genetics , microbiology and biotechnology , gene , embryonic stem cell , embryogenesis , mesoderm
Summary: 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‐ Sce I was coinjected into fertilized eggs with a transgene construct containing flanking recognition sites for I‐ Sce I. 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. genesis 35:248–259, 2003. © 2003 Wiley‐Liss, Inc.