Multiplex genome engineering of livestock stem cells

Supervisors: Simon Lillico and Bruce Whitelaw

Closing Date:  5th January 2022

Genome editing of single targets in livestock has been robustly demonstrated by our group and others. While simultaneous engineering of multiple targets has been achieved in mice through the use of embryonic stem cells, the equivalent has yet to be reported in livestock due in part to the limited proliferative capacity of primary somatic cells. 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.

Burkard C, Opriessnig T, Mileham AJ, Stadejek T, Ait-Ali T, Lillico SG, Whitelaw CBA, Archibald AL (2018) Pigs Lacking the Scavenger Receptor Cysteine-Rich Domain 5 of CD163 Are Resistant to Porcine Reproductive and Respiratory Syndrome Virus 1 Infection. J Virol 92(16):e00415-18

Niu D, Wei HJ, Lin L, George H, Wang T, Lee IH, Zhao HY, Wang Y, Kan Y, Shrock E, Lesha E, Wang G, Luo Y, Qing Y, Jiao D, Zhao H, Zhou X, Wang S, Wei H, Güell M, Church GM, Yang L (2017) Inactivation of porcine endogenous retrovirus in pigs using CRISPR-Cas9.Science 357(6357):1303-1307.

Wang H, Yang H, Shivalila CS, Dawlaty MM, Cheng AW, Zhang F, Jaenisch R (2013) One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering.Cell 153(4):910-8.  

Funding information and application procedures

We would encourage applicants to list up to three projects of interest (ranked 1st, 2nd and 3rd choice) from those listed with a closing date of 5th January 2022 at https://www.ed.ac.uk/roslin/work-study/opportunities/studentships 

This opportunity is open to UK and international students and provides funding to cover stipend, tuition fees and consumable/travel costs.  Applications including a statement of interest and full CV with names and addresses (including email addresses) of two academic referees, should be emailed to [email protected].

When applying for the studentship please state clearly the project title/s and the supervisor/s in your covering letter.

A webinar will be held on Tuesday 14th December at 13.00hrs (UK time) to assist you in the application process and tell you a little more about studying for a PhD with us.  It will also give you an opportunity to ask any questions you may have.  If interested in joining us please send your name and email address to [email protected]by Friday 10th December and we will send you a link to the on line meeting

ALL APPLICATION PROCEDURES MUST BE COMPLETED BY THE CLOSING DATE 5th JANUARY 2022