EASTBIO: Multiplex genome engineering of livestock stem cells at the University of Edinburgh

Supervisors:  Dr Simon Lillico, Professor Bruce Whitelaw.

The genome can be engineered by transgenic and/or genome exiting technologies. For livestock, this has been robustly demonstrated by our group and others for single genomic targets. Although engineering of multiple targets simultaneously has been achieved in mice, this remains largely aspirational for livestock. The challenge is less a question of the efficacy of the engineering tools but more the predominant repair pathway in the target cell. In mice, five genome edits have been achieved simultaneously through the use of murine embryonic stem cells. For livestock the equivalent has yet to be reported; however, multiplexing has been reported for pigs through the simultaneous inactivation of the Porcine Endogenous Retrovirus pol gene at multiple sites in the pig genome. This ‘tour de force’ project tested the boundaries of multiplexing, requiring a cocktail of anti-apoptotic reagents to suppress cell death associated with the resultant genomic instability induced in the pig somatic cells. This strategy of editing all sites in one procedure, while technically challenging, was the only viable approach in primary somatic cells due to their limited proliferative capacity in culture before the onset of senescence. Self-renewing embryonic (or embryonic-like) cells would undoubtedly present an improved platform for multiplexing.

Roslin Technologies are developing pluripotent livestock stem cell resources. This studentship project will use these cells as the basis for developing and demonstrating multiplex genetic engineering (genome editing and/or transgenesis). Gene targets will be selected to clearly show the versatility of engineering protocols and could include engineering of pathways, candidate disease susceptibility genetic loci, gene regulatory loci and transgene ‘landing pads’. Approaches to large scale editing will be investigated, to allow the simultaneous editing of multiple genetic loci associated with polygenic traits. Experiments will demonstrate capability in various species as resources are further developed by Roslin Technologies.

This would be a genuine partnership project. Choice of genetic targets / transgenes will be made jointly, with the genetic engineering tools and reagents made in the Roslin lab and the cell work performed under supervision of the industrial partner.

The student will gain a broad set of skills including the culture and differentiation of pluripotent livestock cells, design and application of both transgenes and CRISPR genome editor reagents, nucleic acid purification and amplification, sequence analysis and western blot. The student will attend weekly research seminars at the Roslin Institute and develop presentation and communication skills through poster and oral presentations at Roslin internal seminars and UK/international meetings.

This 4 year PhD project is part of a competition funded by EASTBIO BBSRC Doctoral Training Partnership (DTP) http://www.eastscotbiodtp.ac.uk/how-apply-0 .

EASTBIO Application and Reference Forms can be downloaded via   http://www.eastscotbiodtp.ac.uk/how-apply-0

Please send your completed EASTBIO Application Form along with a copy of your academic transcripts to [email protected]

You should also ensure that two references have been send to [email protected]  by the deadline using the EASTBIO Reference Form.

This opportunity is open to UK and international students and provides funding covering stipend and UK level tuition fees.  The University of Edinburgh covers the difference between home and international fees meaning that the EASTBIO DTP offers fully-funded studentships to all appointees. 

There is a cap on the number of international students the DTP recruits.  It is therefore important for us to know from the outset which fees status category applicants will fall under when applying to our university.

Please refer to UKRI websiteand Annex B of the UKRI Training Grant Terms and Conditionsfor full eligibility criteria.

Closing Date: 16th December 2021