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Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano
Regeneration-capable flatworms are informative research models to study the mechanisms of stem cell regulation, regeneration, and tissue patterning. However, the lack of transgenesis methods considerably hampers their wider use. Here we report development of a transgenesis method for Macrostomum lig...
Autores principales: | , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5730564/ https://www.ncbi.nlm.nih.gov/pubmed/29242515 http://dx.doi.org/10.1038/s41467-017-02214-8 |
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author | Wudarski, Jakub Simanov, Daniil Ustyantsev, Kirill de Mulder, Katrien Grelling, Margriet Grudniewska, Magda Beltman, Frank Glazenburg, Lisa Demircan, Turan Wunderer, Julia Qi, Weihong Vizoso, Dita B. Weissert, Philipp M. Olivieri, Daniel Mouton, Stijn Guryev, Victor Aboobaker, Aziz Schärer, Lukas Ladurner, Peter Berezikov, Eugene |
author_facet | Wudarski, Jakub Simanov, Daniil Ustyantsev, Kirill de Mulder, Katrien Grelling, Margriet Grudniewska, Magda Beltman, Frank Glazenburg, Lisa Demircan, Turan Wunderer, Julia Qi, Weihong Vizoso, Dita B. Weissert, Philipp M. Olivieri, Daniel Mouton, Stijn Guryev, Victor Aboobaker, Aziz Schärer, Lukas Ladurner, Peter Berezikov, Eugene |
author_sort | Wudarski, Jakub |
collection | PubMed |
description | Regeneration-capable flatworms are informative research models to study the mechanisms of stem cell regulation, regeneration, and tissue patterning. However, the lack of transgenesis methods considerably hampers their wider use. Here we report development of a transgenesis method for Macrostomum lignano, a basal flatworm with excellent regeneration capacity. We demonstrate that microinjection of DNA constructs into fertilized one-cell stage eggs, followed by a low dose of irradiation, frequently results in random integration of the transgene in the genome and its stable transmission through the germline. To facilitate selection of promoter regions for transgenic reporters, we assembled and annotated the M. lignano genome, including genome-wide mapping of transcription start regions, and show its utility by generating multiple stable transgenic lines expressing fluorescent proteins under several tissue-specific promoters. The reported transgenesis method and annotated genome sequence will permit sophisticated genetic studies on stem cells and regeneration using M. lignano as a model organism. |
format | Online Article Text |
id | pubmed-5730564 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-57305642017-12-18 Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano Wudarski, Jakub Simanov, Daniil Ustyantsev, Kirill de Mulder, Katrien Grelling, Margriet Grudniewska, Magda Beltman, Frank Glazenburg, Lisa Demircan, Turan Wunderer, Julia Qi, Weihong Vizoso, Dita B. Weissert, Philipp M. Olivieri, Daniel Mouton, Stijn Guryev, Victor Aboobaker, Aziz Schärer, Lukas Ladurner, Peter Berezikov, Eugene Nat Commun Article Regeneration-capable flatworms are informative research models to study the mechanisms of stem cell regulation, regeneration, and tissue patterning. However, the lack of transgenesis methods considerably hampers their wider use. Here we report development of a transgenesis method for Macrostomum lignano, a basal flatworm with excellent regeneration capacity. We demonstrate that microinjection of DNA constructs into fertilized one-cell stage eggs, followed by a low dose of irradiation, frequently results in random integration of the transgene in the genome and its stable transmission through the germline. To facilitate selection of promoter regions for transgenic reporters, we assembled and annotated the M. lignano genome, including genome-wide mapping of transcription start regions, and show its utility by generating multiple stable transgenic lines expressing fluorescent proteins under several tissue-specific promoters. The reported transgenesis method and annotated genome sequence will permit sophisticated genetic studies on stem cells and regeneration using M. lignano as a model organism. Nature Publishing Group UK 2017-12-14 /pmc/articles/PMC5730564/ /pubmed/29242515 http://dx.doi.org/10.1038/s41467-017-02214-8 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Wudarski, Jakub Simanov, Daniil Ustyantsev, Kirill de Mulder, Katrien Grelling, Margriet Grudniewska, Magda Beltman, Frank Glazenburg, Lisa Demircan, Turan Wunderer, Julia Qi, Weihong Vizoso, Dita B. Weissert, Philipp M. Olivieri, Daniel Mouton, Stijn Guryev, Victor Aboobaker, Aziz Schärer, Lukas Ladurner, Peter Berezikov, Eugene Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano |
title | Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano |
title_full | Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano |
title_fullStr | Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano |
title_full_unstemmed | Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano |
title_short | Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano |
title_sort | efficient transgenesis and annotated genome sequence of the regenerative flatworm model macrostomum lignano |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5730564/ https://www.ncbi.nlm.nih.gov/pubmed/29242515 http://dx.doi.org/10.1038/s41467-017-02214-8 |
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