PhD Study : Permanent gene-based therapy: Application of CRISPR/Cas9 targeted nuclease gene therapy to genetic eye disease

Summary

Background to the project: When the clear window at the front of the eye, known as the cornea, is damaged by injury or disease (including hereditary genetic disease, passed from parents to their children), it can become less transparent causing loss of sight and, at times, blindness. If the cornea becomes too damaged the only treatment available is a corneal transplant from a donor eye. Depending upon which part of the cornea is damaged, all or part of it can be replaced. However, like all organ donations, there is a shortage of donated eyes. Furthermore, as with any intraocular surgery, there are significant inherent risks, which include the potential of a blinding infection, or subsequent rejection or failure of the graft with time. After 10 years almost half of corneal transplants will have failed meaning that the patient will need another transplant. The risk of rejection increases with subsequent transplants. It is possible to remove a small number of corneal epithelial cells from a patient’s eye and grow sufficient numbers of them in the laboratory to transplant them back onto the eye. Our laboratory has shown it is possible to correct serious genetic defects within these cells while they are growing in the laboratory, allowing the patient’s own, now healthy normal cells to be placed back on the eye to restore vision.

Methods to be used: The successful candidate will conduct cutting-edge research in state-of-the art facilities and gain wide experience in a variety of cell and molecular biology techniques. Samples from patients with and without corneal disease and established mouse models of corneal disease are already available. The student will join a group with both national and international collaborations that will allow pursuit of research towards the prevention of this debilitating disease.

Objectives of the Research: CRISPR/Cas9 is a bacterial enzyme that can be used for genome engineering with unprecedented potential for gene therapy. Its advantage over previous technology is that it is easily reprogrammed, by simply changing a short stretch (20 bases) of RNA sequence, to cut DNA at different positions in the genome, depending on the target one wishes to edit. It can be used to knockout a gene, repair an error or direct the delivery of a therapeutic gene to a specified location. This project aims to test the ability of this enzyme to prevent the development of corneal disease using, as a model of corneal disease, mice which have been genetically engineered to develop different forms of the human disease. The ultimate aim of this study is to develop a safe and effective way to get this treatment to the cells that need it in the eye.

Skills required of applicant: The applicant should have experience of undertaking a research project, good communication and organisational skills, experience of effective team working and an ability and willingness to learn new skills and techniques, undertake scientific writing and to travel.

Applications are welcomed from both medical and non- medical graduates.

References:

1.Christie KA et al. 2017. Towards personalised allele-specific CRISPR gene editing to treat autosomal dominant disorders. Sci Rep Nov 22; 7(1):16174. doi:10.1038/s41598-017-16279-4

2.Allen EH, et al. 2016. Keratin 12 missense mutation induces the unfolded protein response and apoptosis in Meesmann epithelial corneal dystrophy. Hum Mol Genet 25:1176.

3.Courtney DG, et al. 2016. CRISPR/Cas9 DNA cleavage at SNP-derived PAM enables both in vitro and in vivo KRT12 mutation-specific targeting. Gene Ther 23:108

4.Courtney DG, et al. 2014. siRNA silencing of the mutant keratin 12 allele in corneal limbal epithelial cells grown from patients with Meesmann's epithelial corneal dystrophy. IOVS 55:3352.

5.Courtney DG, et al. 2014. Development of allele-specific gene-silencing siRNAs for TGFBI Arg124Cys in lattice corneal dystrophy type I. IOVS 55:977.

6.Allen EHA, et al. 2013. Allele-Specific siRNA Silencing for the Common Keratin 12 Founder Mutation in Meesmann Epithelial Corneal Dystrophy. IOVS 54:494.

7.Liao H, et al. 2012. Development of allele-specific therapeutic siRNA in Meesmann epithelial corneal dystrophy. PLoS ONE  6:e28582

8.McLean WHI and Moore CBT. 2011. Keratin disorders – from gene to therapy. Hum Mol Genet 20(R2):R189.

9.Atkinson S, et al. 2011. Development of allele-specific therapeutic siRNA for keratin 5 mutations in epidermolysis bullosa simplex. J Invest Dermatol 131:2079

Essential criteria

Applicants should hold, or expect to obtain, a First or Upper Second Class Honours Degree in a subject relevant to the proposed area of study.

We may also consider applications from those who hold equivalent qualifications, for example, a Lower Second Class Honours Degree plus a Master’s Degree with Distinction.

In exceptional circumstances, the University may consider a portfolio of evidence from applicants who have appropriate professional experience which is equivalent to the learning outcomes of an Honours degree in lieu of academic qualifications.

• Sound understanding of subject area as evidenced by a comprehensive research proposal
• A comprehensive and articulate personal statement

Desirable Criteria

If the University receives a large number of applicants for the project, the following desirable criteria may be applied to shortlist applicants for interview.

• First Class Honours (1st) Degree
• Completion of Masters at a level equivalent to commendation or distinction at Ulster
• Research project completion within taught Masters degree or MRES
• Experience using research methods or other approaches relevant to the subject domain
• Work experience relevant to the proposed project
• Publications - peer-reviewed
• Experience of presentation of research findings

Funding and eligibility

The University offers the following levels of support:

Vice Chancellors Research Studentship (VCRS)

The following scholarship options are available to applicants worldwide:

• Full Award: (full-time tuition fees + £19,000 (tbc))
• Part Award: (full-time tuition fees + £9,500)
• Fees Only Award: (full-time tuition fees)

These scholarships will cover full-time PhD tuition fees for three years (subject to satisfactory academic performance) and will provide a £900 per annum research training support grant (RTSG) to help support the PhD researcher.

Applicants who already hold a doctoral degree or who have been registered on a programme of research leading to the award of a doctoral degree on a full-time basis for more than one year (or part-time equivalent) are NOT eligible to apply for an award.

Please note: you will automatically be entered into the competition for the Full Award, unless you state otherwise in your application.

Department for the Economy (DFE)

The scholarship will cover tuition fees at the Home rate and a maintenance allowance of £19,000 (tbc) per annum for three years (subject to satisfactory academic performance).

This scholarship also comes with £900 per annum for three years as a research training support grant (RTSG) allocation to help support the PhD researcher.

• Candidates with pre-settled or settled status under the EU Settlement Scheme, who also satisfy a three year residency requirement in the UK prior to the start of the course for which a Studentship is held MAY receive a Studentship covering fees and maintenance.
• Republic of Ireland (ROI) nationals who satisfy three years’ residency in the UK prior to the start of the course MAY receive a Studentship covering fees and maintenance (ROI nationals don’t need to have pre-settled or settled status under the EU Settlement Scheme to qualify).
• Other non-ROI EU applicants are ‘International’ are not eligible for this source of funding.
• Applicants who already hold a doctoral degree or who have been registered on a programme of research leading to the award of a doctoral degree on a full-time basis for more than one year (or part-time equivalent) are NOT eligible to apply for an award.

Due consideration should be given to financing your studies. Further information on cost of living