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Characterization of Dengue and Zika viral activities on cellular biology to identify possible strategies to target viral infection or possible uses of those viruses against cancer
University of Costa Rica
Faculty of Microbiology, University of Costa Rica
Research Center on Tropical Diseases, CIET
Dr. Rodrigo Mora Rodríguez, PhD.
English, Spanish and German
Type of Research Project
- Basic science
What is the background of the project?
DENV and ZIKV ( if you use abbreviations please also provide definitions on it, for ex.: PCR(polymeraze chain reacton) are two flaviviruses with an important impact on human health. They are viruses with relatively unstable RNA genomes leading to an enormous variation among the different strains for both viruses. These strains could impact cell biology in different ways leading to variation regarding mainly three factors: 1. High or low permissiveness, 2. High or low viral replication and 3. Survival or Cell Death. The characterization of this diversity has been hampered by the lack of reliable biosensors to report the cellular changes upon infection. We have developed an activable GFP-based sensor that becomes fluorescent upon proteolytic cleavage by DENV and ZIKV proteases, turning the infected cell fluorescent and reporting thereby the presence of viral proteins as surrogate marker of viral replication. Using this system, we quantify and characterize viral infection kinetics by tracking the time-resolved behavior of the fluorescent cells within a Dulbecco Plaque formation assay. Combining this sensor with sensors for autopaghy, apoptosis and necrosis, we will be able to characterize the cellular response to viral replication in terms of programmed cell death subroutines. By means of this caracterization, we envisaged a strategy to identify differences in viral strains to further research on targets to limit viral replication or viral genetic engineering to target cancer cells. Abbreviations: DENV: Dengue virus ZIKV: Zika virus RNA: Ribonucleic acid GFP: Encoding green fluorescent protein
What is the aim of the project?
To characterize the kinetics of viral infection of a strain of DENV or ZIKV and the cellular outcomes in terms of cell death and autophagy.
What techniques and methods are used?
- Culture of cells to infect them with viral strains: Some examples of cell culture include slitting, seeding, counting and others more Staining of the cells with probes for cell death 1- Image analysis using the cellprofiler software Automated microscopy with live imaging of Dulbecco plaque formation assay Image Analysis and interpretation.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
- The intention is that the student can develop microscopic abilities and of the cell culture, and that the student have a closer interaction with the virus, and the opportunity to create his/hers own study cells. - The intention is that the student knows more staining techniques and other techniques for the virus identification. - The intention is that the student gathers his/hers viruses and make his/hers own culture. - The intention is that the student uses his/hers own knowledge to make possible strategies according to the behavior of the virus for its utilization in the cancer. - The intention is that the student analyze, study and learn about different types of DENV and ZIKV that are presented in Costa Rica. - The intention is that the student can recognize and learn to use the cell profiler software.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
1- Training in image analysis using CellProfiler Software (during the internship)
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a poster - The student will prepare a presentation
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
1- Lab training, preferentially cell culture techniques (splitting, seeding, counting)
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system - Students in biomedical fields
- Hamel R; Dejarnac O; Wichit S; Ekchariyawat P; Neyret A; Luplertlop N; Perera-Lecoin M; Surasombatpattana P; Talignani L; Thomas F; Cao-Lormeau V-M; Choumet V; Briant L; Desprès P; Amara A; Yssel H; Missé D. 2015. Biology of Zika virus infection in human skin cells. J Virol 89:8880–8896. doi:10.1128/JVI.00354-15.
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