Developing of a new imaging tool for a single gene locus

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. For example, dCas9 from CRISPR 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 (at least 36 gRNAs) in living human cell [1]. We utilize different tools to study gene locus movement in embryonic stem cells and mammalian cells to discover novel mechanisms of gene regulation.

Bildet kan inneholde: linje, diagram, ledd.

Our lab utilise CRISPR-Cas9 for generegulation and imaging. Illustration by Naima Azouzi.

Project: This master project will be a part of a project funded by Norwegian Research Council working on the characterization of live-cell imaging of individual gene loci. The master student will use various systems and perform transfections followed by imaging. The project is part of a subteam in the lab focusing on development of new imaging techniques to study individual genes within the nucleus of a cell. We use imaging and different knockdown methods (RNAi/CRISPRi/auxin inducible degron) to characterize the mechanistic role of movement of specific gene loci.

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 Chromatome Team (www.chromatome.no). You will learn to make good scientific presentations, have nice 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 (https://www.med.uio.no/cancell/english/) and part of the master project will be at the Medical Faculty, UiO. We can also train you in good scientific writing (https://www.med.uio.no/cancell/english/news-and-events/news/2020/crispr-cas9-article-from-eskeland-lab-in-aftenpost.html).

 

To learn more about our lab visit: https://www.youtube.com/watch?v=6yADYLbFEo4&feature=youtu.be

References:

1.           Chen B, Gilbert LA, Cimini BA, Schnitzbauer J, Zhang W, Li G-W, et al. Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system. Cell. 2013;155: 1479–1491. doi:10.1016/j.cell.2013.12.001

Publisert 6. juli 2020 14:39 - Sist endret 6. juli 2020 14:45

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