This master project is part of a a Centre of Excellence at UiO, the Centre for Cancer Cell Reprogramming.
Background:
In eukaryotes, the genetic information of a cell is packaged into chromatin. The differentiation of embryonic stem cells (ESCs) to all somatic cell types is dependent on changes to gene expression programs defined by the underlying chromatin compaction levels. Measurement of movement of genomic loci is crucial to understand how nuclear positioning and looping of chromatin can regulate gene expression. New tools have been developed which can be used to fluorescently label specific loci in the genome and visualize by live-cell imaging (Wu et al., 2019). For example, dCas9 from theCRISPR system fused to enhanced green fluorescent protein (EGFP) is used to visualise repetitive DNA sequences (telomeres and centromeres) with a single gRNA or non-repetitive loci using multiple gRNAs in living human cell. We utilize different molecular tools to study gene locus movement in embryonic stem cells and mammalian cell lines to discover novel mechanisms of gene regulation.
Project:
This master project will focus on the characterization of live-cell imaging of individual gene loci. The project focus on innovative development of new imaging techniques to study individual genes within the nucleus of a cell (One tool has already a 'Disclosure of Invention'). The master student will use various molecular tools developed in the lab and perform transfections followed by confocal and live-cell imaging. We use imaging and different knockdown methods (RNAi/CRISPRi/degron system) to characterize the mechanistic role of movement of specific gene loci. Furthermore, the student will perform recombinant protein purification and characterization of DNA binding properties of proteins.
Methods:
The student will learn protein purification, mammalian tissue culture including culture of embryonic stem cells, various molecular and immunological methods such as cloning, transfection, western blotting, immunofluoresence and confocal imaging.
About you:
We are looking for a driven, enthusiastic and hardworking student. You should be motivated to learn, have good collaborative skills and be involved in developing new methods. An interest in microscopy is essential.
About us:
During this master project you will be part of our Chromatin Biology Group. You will learn to make good scientific presentations, have creative discussions and will be able to attend national scientific retreats as a part of a young active ambitious research group.
You will be participating in a Centre of Excellence “Centre for Cancer Cell Reprogramming” at UiO and part of the master project will be at the Medical Faculty, UiO. We will also train you in good scientific writing. To learn more about our lab visit: https://www.youtube.com/watch?v=6yADYLbFEo4&feature=youtu.be
Supervisors:
Main supervisor: Ragnhild Eskeland (IMB)
Co-supervisors: Nikolina Seculic (NCMM/IMB)
Internal supervisor: Cinzia Anita Maria Progida (IBV)
References:
Wu, X., Mao, S., Ying, Y., Krueger, C.J., Chen, A.K., 2019. Progress and Challenges for Live-cell Imaging of Genomic Loci Using CRISPR-based Platforms. Genomics Proteomics Bioinformatics. https://doi.org/10.1016/j.gpb.2018.10.001