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2. TALEN-Based Knockout System

Author

Yoshida, Keita, Treen, Nicholas, COHEN, IRUN R., Series Editor, LAJTHA, ABEL, Series Editor, LAMBRIS, JOHN D., Series Editor, PAOLETTI, RODOLFO, Series Editor, REZAEI, NIMA, Series Editor, and Sasakura, Yasunori, editor

Published
2018
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14. GABA-Induced GnRH Release Triggers Chordate Metamorphosis

Author

Hozumi, Akiko, primary, Matsunobu, Shohei, additional, Mita, Kaoru, additional, Treen, Nicholas, additional, Sugihara, Takaho, additional, Horie, Takeo, additional, Sakuma, Tetsushi, additional, Yamamoto, Takashi, additional, Shiraishi, Akira, additional, Hamada, Mayuko, additional, Satoh, Noriyuki, additional, Sakurai, Keisuke, additional, Satake, Honoo, additional, and Sasakura, Yasunori, additional

Published
2020
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26. Mollusc gonadotropin-releasing hormone directly regulates gonadal functions: A primitive endocrine system controlling reproduction

Author

Treen, Nicholas, primary, Itoh, Naoki, additional, Miura, Hanae, additional, Kikuchi, Ippei, additional, Ueda, Takenori, additional, Takahashi, Keisuke G., additional, Ubuka, Takayoshi, additional, Yamamoto, Kazutoshi, additional, Sharp, Peter J., additional, Tsutsui, Kazuyoshi, additional, and Osada, Makoto, additional

Published
2012
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29. Hox-mediated endodermal identity patterns pharyngeal muscle formation in the chordate pharynx.

Author

Yoshida K, Nakahata A, Treen N, Sakuma T, Yamamoto T, and Sasakura Y

Subjects
Animals, Ciona intestinalis, Endoderm drug effects, Feedback, Physiological drug effects, Pharyngeal Muscles drug effects, Pharynx drug effects, Tretinoin pharmacology, Endoderm embryology, Homeodomain Proteins metabolism, Muscle Development drug effects, Pharyngeal Muscles embryology, Pharynx embryology
Abstract

The chordate pharynx, possessing gill slits and the endostyle, is a complex of multiple tissues that are highly organized along the anterior-posterior (AP) axis. Although Hox genes show AP coordinated expression in the pharyngeal endoderm, tissue-specific roles of these factors for establishing the regional identities within this tissue have not been demonstrated. Here, we show that Hox1 is essential for the establishment of AP axial identity of the endostyle, a major structure of the pharyngeal endoderm, in the ascidian Ciona intestinalis We found that knockout of Hox1 causes posterior-to-anterior transformation of the endostyle identity, and that Hox1 represses Otx expression and anterior identity, and vice versa. Furthermore, alteration of the regional identity of the endostyle disrupts the formation of body wall muscles, suggesting that the endodermal axial identity is essential for coordinated pharyngeal development. Our results demonstrate an essential role of Hox genes in establishment of the AP regional identity in the pharyngeal endoderm and reveal crosstalk between endoderm and mesoderm during development of chordate pharynx., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2017. Published by The Company of Biologists Ltd.)

Published
2017
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30. Genome Editing of the Ascidian Ciona intestinalis with TALE Nuclease.

Author

Sasakura Y, Yoshida K, and Treen N

Subjects
Animals, Electroporation, Gene Knockout Techniques, Organ Specificity, Transcription Activator-Like Effector Nucleases metabolism, Ciona intestinalis genetics, Gene Editing methods
Abstract

The ascidian Ciona intestinalis is an important model animal for studying developmental mechanisms for constructing the chordate body. Although molecular and embryological techniques for manipulating Ciona genes were developed a long time ago, recent achievements of genome editing in this animal have innovated functional analyses of genes in Ciona. Particularly, knockout of genes in the G0 generation coupled with tissue-specific expression of TALENs enables us to rapidly address gene functions that were difficult using previous methods.

Published
2017
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31. Germ cell mutations of the ascidian Ciona intestinalis with TALE nucleases.

Author

Yoshida K, Treen N, Hozumi A, Sakuma T, Yamamoto T, and Sasakura Y

Subjects
Animals, Deoxyribonucleases metabolism, Genome, Mutagenesis, Ciona intestinalis enzymology, Ciona intestinalis genetics, Deoxyribonucleases genetics, Electroporation methods, Gene Knockout Techniques methods, Germ Cells metabolism
Abstract

Targeted mutagenesis of genes-of-interest, or gene-knockout, is a powerful method to address the functions of genes. Engineered nucleases have enabled this approach in various organisms because of their ease of use. The ascidian Ciona intestinalis is an excellent organism to analyze gene functions by means of genetic technologies. In our previous study, we reported mutagenesis of Ciona somatic cells with TALE nucleases (TALENs) by electroporating expression constructs. In this study, we report germ cell mutagenesis of Ciona by microinjecting mRNAs encoding TALENs. TALEN mRNAs introduced mutations to target genes in both somatic and germ cells. TALEN-mediated mutations in the germ cell genome were inherited by the next generation. We conclude that knockout lines of Ciona that have disrupted target genes can be established through TALEN-mediated germ cell mutagenesis., (© 2014 Wiley Periodicals, Inc.)

Published
2014
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