Development of genetic manipulation tools in Macrostomum lignano for dissection of molecular mechanisms of regeneration
| PhD ceremony: | Mr J.J. (Jakub) Wudarski |
| When: | January 14, 2019 |
| Start: | 12:45 |
| Supervisor: | prof. dr. E. (Eugene) Berezikov |
| Co-supervisor: | prof. dr. V. (Victor) Guryev |
| Where: | Academy building RUG |
| Faculty: | Medical Sciences / UMCG |
Development of genetic manipulation tools in Macrostomum lignano for dissection of molecular mechanisms of regeneration
The thesis is centered on the development and use of genetic manipulation tools for the flatworm Macrostomum lignano. We present how access to transgenic techniques can facilitate the research in the field of flatworm biology.The introduced molecular toolkit was used to establish a series of transgenic lines that express fluorescent marker protein under the control of tissue specific promoters. We also presented how these lines allowed for FACS isolation and sequencing of flatworm cell populations. The generated RNA-seq data improved the currently existing transcriptomic profiles of neoblasts expanding our knowledge of the stem cell system of the flatworms. An important follow up will be finding neoblast-specific marker genes. A transgenic line expressing fluorescent protein in the stem cells would enable live imaging, lineage tracing and FACS, leading to a more complete characterization of the neoblast system. This could be achieved by either finding a neoblast-specific promoter or tagging a neoblast-specific gene with the help of genome engineering tool such as CRISPR/Cas9.The work presented in this thesis opens a range of future opportunities in the fields of flatworm biology, stem cell research and ageing. One of the important factors to keep in mind is that the method presented is by large a proof-of-principle work. While the delivery method has been refined and optimized, the genetic manipulation tools can still be improved and expanded.