Doctoral Candidate 2
Gene editing for cystic fibrosis – tackling the translatability hurdle
PROJECT INFORMATION
Host institution: KU Leuven, Belgium
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Supervisory team: Prof. Marianne Carlon (PhD promoter, KU Leuven), Prof. Anna Cereseto (Universita degli studi di Trento), Dr. Antonio Casini (Alia Therapeutics)
PROJECT DESCRIPTION
This project is focussed on novel gene editing modalities and delivery methods to develop a gene therapy treatment for the monogenetic disease cystic fibrosis which primarily manifests with lung disease. Objectives: (1) To use gene editing (in particular base and prime editing) for efficient correction of CFTR (gene mutations in CFTR are responsible for cystic fibrosis) in primary human organoid and airway models. (2) To develop safe and efficient delivery vehicles for the gene editing machinery, allowing to ultimately target human lung epithelial cells. (3) To determine the influence of the (diseased) lung micro-environment in gene editing delivery and editing efficiency. The project will involve wet lab work including the design and cloning of DNA constructs, application of viral vectors (adeno-associated viral vectors) and virus like particles (VLPs), working with primary cells and organ-on-chip systems, immunohistochemistry, microscopy, flow cytometry and other biochemical analyses, and molecular biology including bio-informatic evaluation for gene editing efficiency and safety assessment based on sequencing analysis (Sanger, NGS).
A successful project will result in: (1) A new tool to gene edit CFTR mutations which can effectively and safely target the (human) lung. (2) Analysis of the role of the lung microenvironment, in particular inflammation and sputum, which is increased in cystic fibrosis patients, on viral particle delivery (3) Assessment of the role of mucus, inflammation, presence of immune cells, as well as the delivery route, to optimise targeting the desired cells in advanced lung-on-chip models.
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Enrolment in Doctoral School: KU Leuven
Planned secondments:
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INSERM, France (Month 23-24): introduction to the technology platform of virus like particles (‘Nanoblades’)
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Universita degli studi di Trento, Italy (Month 30): introduction to novel nucleases for genome editing
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Alia Therapeutics, Italy (Month 31): Introduction to the VesiCas technology platform
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Mimetas, Netherlands (Month 36-39): application of developed gene editing and delivery technology to Lung on Chip models
ESSENTIAL REQUIREMENTS
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You hold a master’s degree in biochemistry, biotechnology, bio-engineering, biomedical sciences, or a related field.
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You are ambitious, well organized, a team player, and have excellent communication skills.
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You are proficient in English both verbal and written
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You have the ability to work independently and have a critical mindset.
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You are a pro-active and motivated person, eager to participate in network-wide training events, international travel and public awareness activities.
SKILLS AND EXPERTISE
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Solid knowledge of molecular biology and biochemical analyses, and in particular, CRISPR-Cas technology, is an asset. Prior experience working with cell culture, and in particular primary cells, possibly in the field of gene therapy, is considered a plus.