Journal ArticleDOI
Generation of Ci-Brachyury-GFP stable transgenic lines in the ascidian Ciona savignyi.
TLDR
A generation of stable transgenic lines of the ascidian Ciona savignyi carrying a Ciona intestinalis‐Brachyury‐promoter/Green Fluorescent Protein‐reporter (Ci‐Bra‐GFP) construct is reported, indicating integration into the genome.Abstract:
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-SceI was coinjected into fertilized eggs with a transgene construct containing flanking recognition sites for I-SceI. Two founder animals, out of 12 F0 adults tested, were found to transmit the transgene to their offspring (F1s) at frequencies of 42% and 23%. The transgene was further inherited by the F2 in a Mendelian fashion and displayed nonmosaic expression, indicating integration into the genome. The Mendelian inheritance and the absence of mosaicism persisted through the F3 and F4 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.read more
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A muscle-specific transgenic reporter line of the sea anemone, Nematostella vectensis
TL;DR: In this paper, a 1.6-kb fragment of the regulatory upstream region of the Myosin Heavy Chain1 (MyHC1) gene was found to be expressed in the retractor and tentacle muscles of Nematostella polyps, faithfully reproducing the expression of the endogenous MyHC1 gene.
Journal ArticleDOI
Ascidian prickle regulates both mediolateral and anterior-posterior cell polarity of notochord cells.
TL;DR: The ascidian notochord follows a morphogenetic program that includes convergent extension (C/E), followed by anterior-posterior (A/P) elongation, which shows polarity first in the mediolateral (M/L) axis during C/E, and subsequently in the A/P axis during elongation.
Journal ArticleDOI
Transgenesis in fish: efficient selection of transgenic fish by co-injection with a fluorescent reporter construct
TL;DR: A strategy to efficiently generate and preselect transgenic lines harbouring any transgene of interest in medaka and zebrafish by co-injection of a selectable reporter construct and the quality of GFP expression within the F0 generation is described.
Journal ArticleDOI
Ciona intestinalis: chordate development made simple.
TL;DR: The main similarities and differences so far identified, at the molecular level, between ascidian and vertebrate tissues are reviewed and the evolution of the compact ascidian genome is discussed.
Journal ArticleDOI
Piecing together evolution of the vertebrate endocrine system
TL;DR: Analysis of metazoan nucleotide and protein sequences reveals a patchwork pattern of H-P-PG gene conservation between vertebrates and closely related invertebrates (ascidians), consistent with a model of how the vertebrate H-p-PG endocrine system could have emerged in relatively few steps.
References
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Book
Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal ArticleDOI
Repeat-induced gene silencing in mammals.
TL;DR: Use of the loxCre system of site-specific recombination is described to generate transgenic mouse lines in which different numbers of a transgene are present at the same chromosomal location, thereby eliminating the contribution of position effects and allowing analysis of the effect of copy number alone on transGene silencing.
Journal ArticleDOI
Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation
Kristen L. Kroll,Enrique Amaya +1 more
TL;DR: A simple approach for large-scale transgenesis in Xenopus laevis embryos is developed and used to identify in vivo requirements for FGF signaling during gastrulation and, by contrast, embryos expressing XFD contain well-patterned nervous systems despite a putative role for F GF in neural induction.
Journal ArticleDOI
Medaka — a model organism from the far east
TL;DR: In this paper, the authors introduce the use of fish as model genetic systems to study gene function in the context of vertebrate development, using the Zebrafish as a model vertebrate and its Far Eastern cousin Medaka as an important model fish.
Journal ArticleDOI
Cell lineage analysis in ascidian embryos by intracellular injection of a tracer enzyme: III. Up to the tissue restricted stage
TL;DR: Cell lineages during embryogenesis of the ascidian Halocynthia roretzi were analyzed up until the stage where each blastomere was fated to be only a single tissue type by intracellular injection of horseradish peroxidase using the iontophoretic injection method.
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