I have tested a panel of cell-penetrating peptides known to target other intracellular bacteria for their ability to co-localize with Wolbachia within insect cells. Fluorescently-labelled peptides were applied onto Wolbachia-infected cells, their localization observed using a confocal microscope, and co-localization with Wolbachia measured using custom ImageJ macros.
Subsequently, 12 bp synthetic antisense molecules, spanning the ATG and ribosome-binging sites, were designed for two Wolbachia genes: wsp and ftsZ. These genes were selected, as antibodies against them were readily available, and we confirmed their robustness in the silencing protocol. We conjugated the antisense molecules with the cell-penetrating peptides targeting Wolbachia, and applied the onto Wolbachia-infected cells. The knock-down of selected Wolbachia genes was tested with the Western blot using Drosophila tubulin as a loading control. This work, together with the similar protocol using antisense molecules conjugated to cholesterol analogues, has led to the development of the first antisense inhibition protocol for selective and specific targeting of Wolbachia genes.
Although the results of the project were not deployed yet, they were disseminated to both, academic and non-academic audiences in a series of activities. During the project, I participated in the following conferences and scientific events:
1) The British Microbiology Society Annual Conference 2019, Belfast, UK, 08 – 11/04/2019
2) Royal Entomological Society – Special Interest Group: Insects: Infection & Immunity, Online Conference, 24-25 September 2020
3) Virtual Symbiosis Seminar, organized by prof. Greg Hurst and Prof. Nicole Gerardo (
https://docs.google.com/spreadsheets/d/1DHkn2CjRXz9QbJPcSDuibYmn6e1khhgd2xPIxXjXVMY/edit#gid=943392522(öffnet in neuem Fenster)) 7th July 2020
The results of the project have been written up in the following publications/preprints:
- Chrostek E§, Martins NE§, Marialva MS, Teixeira L (2020) Wolbachia-conferred antiviral protection is determined by developmental temperature. bioRxiv, doi: 10.1101/2020.06.24.169169. (citations: 3)
§ Contributed equally
- Corbin C, Jones JE, Chrostek E, Fenton A, Hurst GDD (2020) Thermal sensitivity of the Spiroplasma-Drosophila hydei protective symbiosis: The best of climes, the worst of climes. Molecular Ecology doi: 10.1111/mec.15799.
The approaches and results of the action were disseminated further to non-scientific audience by the fellow invited to act as:
- An Invited author of the article: “Wolbachia – basic science and applications of an antiviral bacterium” for Microbiology Today (
https://bit.ly/3eyZ03T(öffnet in neuem Fenster)) a magazine by the Microbiology Society with articles for all readers, including parliamentarians and policy-makers (06/08/2019).
- An Expert interviewed by “The Scientist Magazine”, a professional magazine providing coverage of recently published scientific papers (
19/07/2019(öffnet in neuem Fenster).
