Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt

Project desciption

Centromeric DNA sequences play an essential role for chromosome distribution in mitosis by recruiting the kinetochore complex that links chromosomes to spindle microtubules. Molecular pathways that influence centromere formation and mitosis directly impact cell division. For instance, during meiosis or in early development, aberrant segregation of chromosomes is a cause of genetic disorders like trisomies, or can lead to developmental arrest and miscarriage. In the adult organism, failure to correctly segregate sister chromatids can lead to neoplastic transformations.
Despite their central function, the molecular mechanisms defining centromere identity and governing centromere assembly remain not well understood. A potential role for an RNA component has been suggested, but the molecular mechanisms and their implications behind, remain unknown. This project aims towards uncovering the functions of centromeric RNA (cenRNA) transcription for centromere assembly and elucidate potential roles of cenRNA in mitotic spindle formation and centromere establishment. 
To identify, track, and investigate associations of cenRNA with centromeric proteins during the cell cycle, the aim is to develop CRISPR-based probes for live-cell RNA tracking and live-cell confocal microscopy. From multidimensional imaging data sets, and molecular as well as pharmacological perturbation experiments in mouse embryonic stem cells, the goal is to obtain a comprehensive picture of the life-cycle of cenRNAs and uncover their functional role in centromere establishment and mitosis. 
As incorrect distribution of chromosomes is linked with hereditary disorders or neoplastic transformation in the case of tumor formation, we expect that understanding how cenRNA impacts chromosome segregation will contribute to insights into the mechanisms by which mitosis progresses normally and how to possibly target, or ameliorate, faulty mitosis in disease.
This project offers an exciting opportunity to work on the interface of RNA biology, CRISPR technology, molecular biology, live-cell confocal microscopy, and image analysis.

Related literature:

O. O. Abudayyeh, et al. (2017), RNA targeting with CRISPR-Cas13. Nature 550, pages 280–284

A. W. Grenfell, R. Heald, M. Strzelecka(2016), Mitotic noncoding RNA processing promotes kinetochore and spindle assembly in Xenopus. The Journal of Cell Biology 214, 133-141

D. Quénet, Y. Dalal (2014), A long non-coding RNA is required for targeting centromeric protein A to the human centromere. eLife 3, 267

Ettinger, A. & Wittmann (2014), T. Fluorescence live-cell imaging. Methods in Cell Biology 123, 77–94