Project properties
Title Pimp that wasp: Nasonia vitripennis genetic engineering for better biocontrol
Group Entomology, Laboratory of
Project type thesis
Credits 36
Supervisor(s) Filippo Guerra, Eveline Verhulst
Examiner(s) Marcel Dicke
Contact info
Begin date 2020/02/28
End date 2024/02/28
Description In the molecular lab at entomology we use for our studies a rather unusual model organism, the parasitoid wasp Nasonia vitripennis. We focus on the use of Nasonia to investigate sex determination, memory development and insect-bacteria interaction to improve biological control agents, while many other research groups use Nasonia for evolutionary biology, development, ecology, and behavior studies. To explore Nasonia biology we can rely on an expanding array of genomic, transcriptomic and functional resources.
ZFN and CRISPR-Cas site directed mutagenesis have been achieved, but the protocol relies on difficult microinjections of the embryos (which are 50x10x10�m big). The small size causes even the most experienced researcher to kill most of the embryos in the process.
We aim to engineer Nasonia without embryonic microinjection thanks to a technique named Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control). To perform ReMOT Control, Cas9 is conjugated to a small peptide (P2C) that mediates its transfer from the hemolymph of the mother to the oocytes. Injecting the mother abdomen it is possible to mutate the offspring. Optimization of ReMOT Control in Nasonia vitripennis would be a major improvement that would avoid embryo microinjection, which can cause death rates of 99.6%.
We will use ReMOT to disrupt DNA-protein interaction and manipulate sex differentiation in Nasonia.

Want to join me in this investigative journey? Then get ready for these challenges:
1) Create plasmids by cloning (Gibson assembly and restriction cloning) to produce heterologous proteins;
2) Inject proteins (eGFP) in Nasonia and confirm localization in the ovaries via confocal microscopy;
3) Inject Cas9 in Nasonia and confirm protein activity in vivo.

If you want to work in a friendly environment and do real molecular science with a twist, contact me at filippo.guerra@wur.nl
Used skills Cloning, heterologous protein production, pupal injection, confocal microscopy, CRISPR-Cas genetic engineering
Requirements For doing a BSc-thesis at Entomology, there are no requirements for specific courses.
For doing a MSc-thesis or internship at Entomology, the following requirements apply: ENT-30806 + a second ENT-course (preferably ENT-30306 or ENT-50806 or ENT-53806). As an alternative for the second ENT-course, PHP-30806 or BHE-30306 can be selected.
Note: these requirements do not apply for MBI students; MBI students should check the requirements for doing an ENT MSc-thesis or internship in the study programme of their specialisation.