Cookies on this website

We use cookies to ensure that we give you the best experience on our website. If you click 'Accept all cookies' we'll assume that you are happy to receive all cookies and you won't see this message again. If you click 'Reject all non-essential cookies' only necessary cookies providing core functionality such as security, network management, and accessibility will be enabled. Click 'Find out more' for information on how to change your cookie settings.

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.

Original publication

DOI

10.1038/s41467-017-02214-8

Type

Journal article

Journal

Nat Commun

Publication Date

14/12/2017

Volume

8

Keywords

Animals, Animals, Genetically Modified, Embryo, Nonmammalian, Female, Gene Expression Profiling, Gene Expression Regulation, Developmental, Gene Transfer Techniques, Genome, Helminth, Luminescent Proteins, Male, Organ Specificity, Ovary, Platyhelminths, Promoter Regions, Genetic, Regeneration, Testis, Transgenes