PhD Study : Engineering plasmid nanoparticles as a platform for delivery of therapeutics

Summary

Background Plasmids have been used to deliver therapeutics in the form of peptides and proteins. As with any other DNA therapeutic, they have low toxicity and are easy to manufacture, store and transport. DNA therapeutics have demonstrated no dose limiting toxicity and do not stimulate production of neutralising antibodies. Thus they can be safely administered repeatedly to achieve a therapeutic endpoint. Unfortunately the uptake of naked DNA plasmids tends to be poor, and they can be rapidly degraded by nucleases in vivo.

Underlying aim

This project seeks to explore the potential of nanosize assembled plasmids as active delivery agents of peptides and proteins. More particularly, the project will explore the potential of assembling double stranded DNA of plasmids into nanostructures with optimized dimensions functionalized for targeted uptake by cells, or tissues for the production of encoded peptides or/and proteins.

Specific objectives/Methodology

1.Design of nanoplasmids. Several designs of nanoplasmids will be performed using currently available software (e.g. CadNano). For proof of concept the designs will be enabled on a plasmid coding for a fluorescent protein. It will also be labelled with fluorescent tags for assessing tissue delivery and intracellular fate.

2.Engineering of nanoplasmids. Common DNA nanotechnology self-assembly strategies will be utilized to engineer the nanostructures, and improve the yields of the nanoplasmid self-assembly.

3.Nanoplasmid characterisation. Characterisation will involve determining the morphology and dimensions utilizing AFM, as well as response to enzymatic degradation.

4.Functionality of nanoplasmid. Tissue culture will be used to evaluate cell targeting, triggering and functionality, of intracellularly delivered nanoplasmids utilizing cell imaging.

5.Target delivery of nanoplasmids. Biodistribution, fate, and triggering of expression of the nanoplasmids will be examined using whole body imaging of animal models.

Requisite experience Applicants will be expected to have cell culture and animal work experience.

References

1.Folding DNA to create nanoscale shapes and patterns. Rothemund PWK. Nature 440(2006)297-302.

2.Encoding DNA quadruplex structure. Dvorkin et al. Science Advances 4(2018)eaat3007.

3.Development of a DNA vaccine targeting human papillomavirus type 16 oncoprotein E6. Peng SW et al. Journal of Virology 16(2004) 8468-8476.

4.Room-temperature and selective triggering of supramolecular assembly-disassembly by non-ionizing radiation. Greschner A.A et al. Journal of the American Chemical Society 141(2019)3456-3469.

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

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.

• Completion of Masters at a level equivalent to commendation or distinction at Ulster
• Experience using research methods or other approaches relevant to the subject domain
• Work experience relevant to the proposed project
• Publications - peer-reviewed
• Publications record appropriate to career stage
• Experience of presentation of research findings
• A comprehensive and articulate personal statement
• Use of personal initiative as evidenced by record of work above that normally expected at career stage.
• Relevant professional qualification and/or a Degree in a Health or Health related area

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