Beltman, F. | Search bioRxiv

bioRxiv2020

Tim29 is required for stem cell activity during regeneration in the flatworm Macrostomum lignano

Domain of Unknown Function (DUF) genes are broadly conserved in metazoa but have an unknown biological function. Previously, we established transcriptional profiles of the germline and stem cells of the flatworm Macrostomum lignano (Grudniewska et al., 2016). Here, we used this information to probe the function of DUF genes in M. lignano. Recently, Kang et al. (2016) found that DUF2366 is a mitochondrial inner membrane protein that interacts with the protein import complex TIM22. DUF2366 was renamed TIM29, and shown to stabilize the TIM22 complex, but its biological role remained largely unknown. We now demonstrate that DUF2366/TIM29 is dispensable in the homeostatic condition in M. lignano, but is essential to adapt to a highly proliferative state required for regeneration. Our results show that de-DUFing the function of DUF genes might require special biological conditions, such as regeneration, and establishes M. lignano as an informative platform to study DUF genes.

cell biology↗

bioRxiv2017

A platform for efficient transgenesis in Macrostomum lignano, a flatworm model organism for stem cell research

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.

bioengineering↗