PhD students in the Laboratory of Chronobiology within the Horizon Europe

Entomologický ústav

Pracovní smlouva

Two PhD projects are available:

• DC1: Non-Drosophila insect neuropeptides: their role in reproduction and seasonality

Objectives: To study neuropeptides lost in ‘higher’ Diptera such as Drosophila and define their roles in circadian, seasonal and life cycle phenotypes. We will take advantage of our recent genomic and transcriptomic data from the linden bug (Pyrrhocoris apterus), from which we have identified neuropeptide candidates present in basal insect lineages but lost in the fruit fly. We will further identify and validate these Heteroptera-specific neuropeptides. We will localize these candidates in brains using in situ hybridization and/or immunohistochemistry (in collaboration with Pyza, JU). Any role in major physiological processes, such as circadian rhythms, reproduction, diapause, mating and the life cycle will be tested using well-established RNA interference. We foresee in-depth analysis of two to three such neuropeptides with potential to reveal important novel insight into insect circadian and seasonal physiology.
Host supervisor: Dr. David Dolezel, co-supervisor: Dr. Vlastimil Smykal.
Planned secondment(s): UNI KASSEL, Prof. Monika Stengl, to learn techniques for primary cell cultures; Jagiellonian University, Prof. Ela Pyza, to perform immunohistochemical experiments.

• DC2: New and optimized reverse genetic tools for diapause research in non-model insects 

Objectives: To develop more efficient methods for gene silencing and gene overexpression in insects. While CRISPR/Cas9 is an extremely versatile method for gene modification that should in principle work in all organisms, the efficient delivery of gRNA/Cas9 complex is species-specific and requires species-tailored protocols for embryo microinjection. Therefore, many groups of organisms, including insect pests and species established as models in physiology, are not practically accessible to reverse genetics. To solve this problem, we will test and optimize recently developed maternal gRNA/Cas9 delivery methods in the linden bug, where gene editing by embryonic gRNA/Cas9 delivery resulted in a robust phenotype. In a parallel research direction, we will explore the use of viral systems, i.e., Negev virus species that we have recently identified in the linden bug and test its expression efficiency using EGFP. The next steps will address if the system, primarily tested on the linden bugs, is accessible to selected species of our consortium (pea aphids, bumblebees, BSF, Musca). Finally, we will overexpress either dominant-negative forms of clock proteins or dsRNA in selected species and test the involvement of the circadian clock in diapause regulation and seasonality.  
Host supervisor: Dr. Vlastimil Smykal, co-supervisor: Dr. David Dolezel.
Planned secondment(s): University Valencia, Prof. Martines-Torres to learn aphid biology, University of Leicester, Prof Rosato, to learn the biology of the black soldier fly.
 

Requirements

• Motivation 
• English at B2 level, written and spoken
• The mobility rule applies: at the time of the recruitment, researchers must not have resided or carried out their main activity (work, studies, etc.) in the Czech Republic for more than twelve months in the three years immediately before the reference date
   

Examples of recent publications from the laboratory of Chronobiology
Tumova S., Dolezel D., Jindra M. (2024) Conserved and Unique Roles of bHLH-PAS Transcription Factors in Insects - From Clock to Hormone Reception. Journal of Molecular Biology 436:1 doi.org/10.1016/j.jmb.2023.168332

Smykal V, Chodakova L, Hejnikova M, Briedikova K, Wu BC, Vaneckova H, Chen P, Janovska A, Kyjakova P, Vacha M, Dolezel D. (2023) Steroid Receptor Coactivator TAIMAN Is a New Modulator of Insect Circadian Clock, PloS Genetics, https://doi.org/10.1371/journal.pgen.1010924 

Smýkal V., Doležel D. (2023) Evolution of proteins involved in the final steps of juvenile hormone synthesis.  J. Insect Physiol 145: March 2023, 104487. DOI: 10.1016/j.jinsphys.2023.104487

Doležel D. (2023) Chapter Molecular Mechanism of the Circadian Clock in Insect Chronobiology, editors Tomioka, Numata, ISBN: 978-981-99-0725-0 Springer Nature

Kotwica-Rolinska J., Chodáková L., Smýkal V., Damulewicz M., Provazník J., Wu C., Hejníková M., Chvalová D., Doležel D. (2022) Loss of Timeless Underlies an Evolutionary Transition within the Circadian Clock. Molecular Biology and Evolution 39(1): msab346 doi: 10.1093/molbev/msab346

Hejníková M., Nouzová M., Ramirez C.E., Fernandez-Lima F., Noriega F., Doležel D. (2022) Sexual dimorphism of diapause regulation in the hemipteran bug Pyrrhocoris apterus. Insect Biochemistry and Molecular Biology 142: 103721. DOI: 10.1016/j.ibmb.2022.103721

Smýkal V., Pivarči M., Provazník J., Bazalová O., Jedlička P., Lukšan O., Horák A., Vaněčková H., Beneš V., Fiala I., Hanus R., Doležel D. (2020) Complex evolution of insect insulin receptors and homologous decoy receptors, and functional significance of their multiplicity. Molecular Biology and Evolution 37: 1775–1789. DOI: 10.1093/molbev/msaa048

Kotwica-Rolinska J., Krištofová L., Chvalová D., Pauchová L., Provazník J., Hejníková M., Sehadová H., Lichý M., Vaněčková H., Doležel D. (2020) Functional analysis and localisation of a thyrotropin-releasing hormone-type neuropeptide (EFLa) in hemipteran insects. Insect Biochemistry and Molecular Biology 122: article number: 103376.: DOI: 10.1016/j.ibmb.2020.103376
 

Our selection procedure is open, transparent, merit-based and in line with the Code of Conduct for the Recruitment of Researchers. 
Candidates should submit the following documents as one single PDF file via the REPLY button below by 15 October 2024 23:59 CET: 

• a detailed CV
• a motivation letter 
• two letters of reference 
• transcripts of bachelor’s and master’s degrees 
• an English certificate

Alternatively, you can send requested documents to incite@entu.cas.cz.

We will choose the candidates that fit the most for the different projects through the following steps:

• Eligibility check: our recruitment committee will check that each application is complete and that applicants fulfill the eligibility criteria described in the previous section
• Remote Evaluation: each eligible application will be assessed independently by the principal investigators of our network, according to the project interests indicated by the applicants. The principal investigators will select up to four candidates per position to pass through to the following stage
• Online interviews: short-listed candidates will be invited for an online interview. Those who have been positively evaluated but not initially called for interviews will be put on a reserve list. Short-listed candidates will be interviewed by their putative supervisor (i.e. the supervisor responsible for the research project of interest), two additional supervisors from the network, and a member of the recruitment committee
• Notification of the selection outcomes: our recruitment committee will notify short-listed candidates of the outcome, and those selected will be put in touch with the corresponding principal investigator and the HR Department of the hosting institution to initiate the hiring procedure. Selected candidates will have to apply to individual PhD programs at host institutes (e.g., applicants interested in the EMBL position will also have to apply to the International EMBL PhD Programme) 

Applicants who have not been successful but received a positive evaluation will be put on a waiting list to cover possible withdrawals and future positions. 

Recruitment Calendar:

• Call opening: September 2024  
• Interviews: September – November 2024 
• Start date of the fellowship: January 2025 – March 2025 (depending on the position and university)