Doctoral Candidate - Gene Editing for Cystic Fibrosis

Gene Editing for Cystic Fibrosis – Tackling the Translatability Hurdle

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:

• 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
• To develop safe and efficient delivery vehicles for the gene editing machinery, allowing to ultimately target human lung epithelial cells
• 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:

• A new tool to gene edit CFTR mutations which can effectively and safely target the (human) lung
• Analysis of the role of the lung microenvironment, in particular inflammation and sputum, which is increased in cystic fibrosis patients, on viral particle delivery
• 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

Enrolment in Doctoral School: KU Leuven

Planned secondments:

1. INSERM, France (Month 23-24): introduction to the technology platform of virus like particles (‘Nanoblades’)
2. Universita degli studi di Trento, Italy (Month 30): introduction to novel nucleases for genome editing;
3. Alia Therapeutics, Italy (Month 31): Introduction to the VesiCas technology platform
4. Mimetas, Netherlands (Month 36-39): application of developed gene editing and delivery technology to Lung on Chip models

Essential requirements of a successful candidate:

• You hold a master’s degree in biochemistry, biotechnology, bio-engineering, biomedical sciences, or a related field
• You are ambitious, well organized, a team player, and have excellent communication skills
• You are proficient in English both verbal and written *You have the ability to work independently and have a critical mindset
• You are a pro-active and motivated person, eager to participate in network-wide training events, international travel and public awareness activities

Skills that are viewed as an extra benefit:

• 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

Contact Person
Marianne Carlon, marianne.carlon@kuleuven.be

Project Description - APPLYMCSA Doctoral Network GET-IN