RT Journal Article
SR Electronic
T1 A platform for efficient transgenesis in <em>Macrostomum lignano</em>, a flatworm model organism for stem cell research
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 151654
DO 10.1101/151654
A1 Jakub Wudarski
A1 Daniil Simanov
A1 Kirill Ustyantsev
A1 Katrien de Mulder
A1 Margriet Grelling
A1 Magda Grudniewska
A1 Frank Beltman
A1 Lisa Glazenburg
A1 Turan Demircan
A1 Julia Wunderer
A1 Weihong Qi
A1 Dita B. Vizoso
A1 Philipp M. Weissert
A1 Daniel Olivieri
A1 Stijn Mouton
A1 Victor Guryev
A1 Aziz Aboobaker
A1 Lukas Schärer
A1 Peter Ladurner
A1 Eugene Berezikov
YR 2017
UL http://biorxiv.org/content/early/2017/06/18/151654.abstract
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 significantly 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 showed 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.