Department Pathology
Principal investigator Martin van Royen
E-mail address –
Website https://www.erasmusmc.nl/en/research/researchers/royen-martin#5d7365b2-5e29-4090-9288-516239906d71
Deciphering the importance of the Glucocorticoid receptor interactome by automated high-throughput FRAP microscopy screens
Suitable as a BEP? No
Suitable as a MEP? Yes
Suitable as an Academic Research Project? No
Techniques:
- Cell culture and molecular cloning
- Generating genetically modified cell lines using CRISPR-Cas9 technology
- Developing and implementation of automated confocal microscopy platform
- Fluorescence imaging data analysis (FRAP)
- Further investigation on functional role of selected cofactors (colocalization analysis, regulation of target genes)
- Literature research and writing a scientific report or thesis
The aim of this student project is to develop and implement a fully automated microscopy pipeline to identify GR cofactors that influence GR-mediated transcription. We will use A549 cells with EGFP-tagged endogenous GR and an MMTV DsRed reporter, stimulating with dexamethasone. A CRISPR screen will be implemented to generate knockout sublines; hits showing altered GR mobility or transcriptional activity will advance to super-resolution colocalization and orthogonal assays, including RT-qPCR, RNA-seq, and RIME. The student will handle cell culture, cloning, CRISPR engineering, automated microscopy pipeline development, image analysis, statistics, data management and reporting including literature review and thesis writing. Daily supervision: Selçuk Yavuz (postdoc), Supervision Dr. Martin E. van Royen and Prof. Dr. Adriaan B. Houtsmuller, Department of Pathology, Erasmus MC.
Further reading (click to link to article)
https://academic.oup.com/nar/article/51/20/10992/7288829?login=true