This master project is part of the Centre for Cancer Cell Reprogramming a Centre of Excellence at UiO (https://www.med.uio.no/cancell/english/). To learn more about us visit www.chromatome.no and watch our movie: https://youtu.be/6yADYLbFEo4
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 regulatory elements located in a large distance from the gene promoter is crucial to understand how nuclear positioning and looping of chromatin can regulate gene expression. Tools have been developed which can be used to fluorescently label specific loci in the genome and visualize by live-cell imaging. For example, dCas9 from CRISPR system fused to enhanced green fluorescent protein (EGFP) is used to visualise repetitive DNA sequences (telomeres and centromeric) with a single gRNA or non-repetitive loci using multiple gRNAs in living human cells. We utilize different tools to study gene locus movement in embryonic stem cells and mammalian cells to discover novel mechanisms of gene regulation.
Project
This master project will be working on the characterization of live-cell imaging of individual gene loci. The master student will use various cell line models and perform transfections followed by confocal imaging. The master student will be part of a sub team in the lab focusing on development of new imaging techniques to study individual genes within the nucleus of a cell.
Methods
The student will learn mammalian tissue culture including culture of embryonic stem cells, various molecular and immunological methods such as molecular cloning, transfection, CRISPR/Cas9 genome editing, Western blotting, Fluorescence in situ hybridization (FISH), immunofluorescence, immunoprecipitation and live-cell and confocal imaging.
About you
We are looking for a driven, enthusiastic and hardworking student. You should be motivated to work in an interdisciplinary environment at IMB. An interest in microscopy is essential.
About us
During this master project you will be part of Chromatin Biology Group (www.chromatome.no). You will learn how to make good scientific presentations, work in a collaborative team and present your work on national scientific retreats as a part of a young active ambitious research group.
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