TY - JOUR
T1 - Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano
AU - Wudarski, Jakub
AU - Simanov, Daniil
AU - Ustyantsev, Kirill
AU - De Mulder, Katrien
AU - Grelling, Margriet
AU - Grudniewska, Magda
AU - Beltman, Frank
AU - Glazenburg, Lisa
AU - Demircan, Turan
AU - Wunderer, Julia
AU - Qi, Weihong
AU - Vizoso, Dita B.
AU - Weissert, Philipp M.
AU - Olivieri, Daniel
AU - Mouton, Stijn
AU - Guryev, Victor
AU - Aboobaker, Aziz
AU - Schärer, Lukas
AU - Ladurner, Peter
AU - Berezikov, Eugene
N1 - Funding Information:
We thank H. Clevers for the support on the early stages of the project; E. Cuppen, E. de Bruin, P. van Zon and H. Lunstroo for the help with generating 454 data, and ERIBA sequencing facility for generating Illumina data. This work was supported by the European Research Council (ERC Starting Grant “MacModel”, grant no. 310765) to E.B., K.U. was supported by the project 0324-2016-0008 from the Russian State Budget. A.A. was supported by the Biotechnology and Biological Sciences Research Council (BBSRC, grant no. BB/K007564/1). P.L. was supported by the Austrian Science Fund (FWF, grant no. 25404). L.S. was supported by the Swiss National Science Foundation (SNFS, grant no. 3100A0-127503 and 31003A-143732). The work on annotation of transposable elements was supported by the Russian Foundation for Basic Research (RFBR, grant no. 15-04-08003) to E.B.
Publisher Copyright:
© 2017 The Author(s).
PY - 2017/12/1
Y1 - 2017/12/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85038815617&partnerID=8YFLogxK
U2 - 10.1038/s41467-017-02214-8
DO - 10.1038/s41467-017-02214-8
M3 - Article
AN - SCOPUS:85038815617
SN - 2041-1723
VL - 8
JO - Nature Communications [E]
JF - Nature Communications [E]
IS - 1
M1 - 2120
ER -