Getting ready for HGP-Write: constructing and shuttling yeast artificial chromosomes from yeast to mammalian cells through cell fusion

This is a UK Centre for Mammalian Synthetic Biology Studentship.

Interested individuals must follow Steps 1, 2 and 3 at this link on how to apply:

http://www.ed.ac.uk/biology/prospective-students/postgraduate/pgr/how-to-apply

Supervisors: Dr Patrick Cai ([Email Address Removed]), Prof Wendy Bickmore ([Email Address Removed]) and Prof Steven Pollard ([Email Address Removed])

Yeast is a great chassis for constructing large artificial chromosomes (> 1Mb) due to its very efficient homologous recombination mechanisms. Through the years, our labs have pioneered technologies to design, synthesis, assemble and integrate artificial yeast chromosomes. On the other hand, even though long range gene regulation is essential for engineering mammalian cells, we lack a robust method to effectively construct large synthetic chromosomes in mammalian cells. In this project, we propose to develop a robust pipeline to construct artificial chromosomes in yeast, and then transfer the artificial chromosomes into embryonic stem cells through ES cell – yeast spheroplast fusion. Through the development of this powerful technology platform, we will study and re-engineer the complex regulatory landscape of mammalian developmental genes. The Pax6 gene encodes a transcription factor involved in development of the brain, eye and pancreas. Pax6 expression at these developmental sites is controlled by a complex set of enhancers scattered over a large (>200kb) genomic interval surrounding Pax6 – including some embedded in introns of other genes. The importance of the genomic location of these enhancers, relative to the gene that they control is not understood. In this project, this question will be addressed by re-engineering the 420kb regulatory landscape of the human PAX6 gene carried in a yeast artificial chromosome. The functionality of this re-written part of the human genome will be tested by functional complementation of mouse cells that are null for Pax6. The human PAX6 YAC has previously been shown to rescue the mutant phenotype of mouse Pax6 mutants.

First supervisor: Dr. Patrick Cai (http://www.cailab.org)

Second supervisor: Prof. Wendy Bickmore (http://www.hgu.mrc.ac.uk/people/bickmore_researchb.html) and Dr. Steve Pollard (http://www.crm.ed.ac.uk/research/group/neural-stem-cells-and-brain-cancer

The PhD student will become part of a cohort of graduates students linked to the research of the new UK Centre for Mammalian Synthetic Biology (the ‘Centre’), based at the University of Edinburgh. Through support from the Research Council’s Synthetic Biology for Growth programme and of the BBSRC, EPSRC and MRC, the University has been awarded ~ £18M in funding to establish a national facility for DNA synthesis (the Edinburgh Genome Foundry) and one of six UK Centres of Excellence in Synthetic Biology. Edinburgh’s Centre embeds colleagues from the College of Medicine and Veterinary Medicine, in particular from the Scottish Centre for Regenerative Medicine. More information about our Centre can be found at http://www.synbio.ed.ac.uk and follow us on Twitter @SynthSysEd.

This is an exciting opportunity to be at the cutting edge of this fast moving area of science and technology in world-leading research institutes. We are looking for highly motivated graduates who are enthusiastic about the potential of this new area of science and keen to work across disciplines.