An enhancer trap in the ascidian Ciona intestinalis identifies enhancers of its Musashi orthologous gene

Awazu, Satoko; Sasaki, Akane; Matsuoka, Terumi; Satoh, Nori; Sasakura, Yasunori

doi:10.1016/j.ydbio.2004.08.029

Cited by 45 publications

(49 citation statements)

References 38 publications

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“…The sj mutant was obtained by Minos-based insertional mutagenesis (10)(11)(12)(13)(14). Our Minos construct contains gfp as a reporter gene (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The PCR-amplified transcription termination sequence of hsp 70 of Drosophila melanogaster (derived from pGaTB) was ligated as a PstI fragment in the PstI site of pMiFr3dTPO-gfp (12) to generate pMiTFr3dTPO-gfp.…”

Section: Methodsmentioning

confidence: 99%

“…In an ascidian, C. savignyi, mutagenesis with a chemical mutagen N-ethyl-N-nitrosourea, successfully revealed the function of prickle (7) and tyrosinase (8). Recently, germ-line transgenesis has been achieved in two ascidians, C. intestinalis and C. savignyi, with a Tc1͞mariner superfamily transposon Minos (9)(10)(11)(12)(13)(14). The insertion of Minos into the Ciona genome indicates that Minos can be used as a mutagen, because Minos disrupts the gene function if it is inserted into a genomic region that encodes a gene.…”

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confidence: 99%

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Transposon-mediated insertional mutagenesis revealed the functions of animal cellulose synthase in the ascidian Ciona intestinalis

Sasakura

Nakashima²,

Awazu³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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Tunicates are the only animals that perform cellulose biosynthesis. The tunicate gene for cellulose synthase, Ci-CesA, was likely acquired by horizontal transfer from bacteria and was a key innovation in the evolution of tunicates. Transposon-based mutagenesis in an ascidian, Ciona intestinalis, has generated a mutant, swimming juvenile (sj). Ci-CesA is the gene responsible for the sj mutant, in which a drastic reduction in cellulose was observed in the tunic. Furthermore, during metamorphosis, which in ascidians convert the vertebrate-like larva into a sessile filter feeder, sj showed abnormalities in the order of metamorphic events. In normal larvae, the metamorphic events in the trunk region are initiated after tail resorption. In contrast, sj mutant larvae initiated the metamorphic events in the trunk without tail resorption. Thus, sj larvae show a ''swimming juvenile'' phenotype, the juvenile-like trunk structure with a complete tail and the ability to swim. It is likely that ascidian cellulose synthase is required for the coordination of the metamorphic events in the trunk and tail in addition to cellulose biosynthesis.Minos ͉ swimming juvenile ͉ Ci-CesA ͉ metamorphosis T unicates are the only animals able to perform cellulose biosynthesis (1-3). Cellulose synthase is an enzyme that has a central role in cellulose biosynthesis; the gene (Ci-CesA) for cellulose synthase is encoded in the Ciona intestinalis genome and expressed in the larval ectoderm (4). Recent molecular phylogenetic analyses of Ci-CesA and Cs-CesA of Ciona savignyi suggest that the gene has been acquired by horizontal transfer from bacteria (5, 6). Ascidian cellulose synthase may be involved in the formation of the tunic, the cellulose-containing structure that surrounds the surface of the body to protect against predators. However, functional analysis showing that ascidian cellulose synthase contributes to the tunic formation in vivo has not been reported.Mutant analysis is a powerful way to understand gene function. In an ascidian, C. savignyi, mutagenesis with a chemical mutagen N-ethyl-N-nitrosourea, successfully revealed the function of prickle (7) and tyrosinase (8). Recently, germ-line transgenesis has been achieved in two ascidians, C. intestinalis and C. savignyi, with a Tc1͞mariner superfamily transposon Minos (9-14). The insertion of Minos into the Ciona genome indicates that Minos can be used as a mutagen, because Minos disrupts the gene function if it is inserted into a genomic region that encodes a gene.Here, we have isolated a mutant, swimming juvenile (sj), which was caused by an insertion of Minos into the C. intestinalis genome. sj shows defects in the tunic structure of swimming larvae. In addition to the tunic phenotype, they showed an abnormal order of metamorphic events, suggesting that the regulation of the timing of these events is affected in sj mutants. Ci-CesA is the gene responsible for the sj mutant, and the drastic loss of cellulose was observed in the tunic of sj mutants. These results indicate that ascidian ...

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“…The sj mutant was obtained by Minos-based insertional mutagenesis (10)(11)(12)(13)(14). Our Minos construct contains gfp as a reporter gene (Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Transposon-mediated insertional mutagenesis revealed the functions of animal cellulose synthase in the ascidian Ciona intestinalis

Sasakura

Nakashima²,

Awazu³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

109

112

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show abstract

“…Microinjection-mediated enhancer detection is the simplest method (Sasakura et al, 2003;Awazu et al, 2004Awazu et al, , 2007. Its technical advantage is that we have only to prepare a transposon vector for enhancer detection to perform this method.…”

Section: Comparison Of the Methods Of Enhancer Detection In Cionamentioning

confidence: 99%

“…In fact, several Tc1/mariner transposons have shown their activity in both protostomes and deuterostomes (Loukeris et al, 1995a,b;Klinakis et al, 2000a, b;Shimizu et al, 2000;Zhang et al, 2002;Drabek et al, 2003;Pavlopoulos and Averof, 2005;Metaxakis et al, 2005). Genetic techniques such as germline transgenesis, enhancer detection (also called "enhancer trapping"), and insertional mutagenesis have been established in Ciona with Minos (Sasakura et al, 2003bAwazu et al, 2004).…”

Section: Introductionmentioning

confidence: 99%