Over the last decade, CRISPR/Cas9-based techniques have been developed for various genomic engineering strategies in eukaryotic cells. Using single guide RNAs (sgRNA) containing specific targeting sequences, Cas9 can target precise genomic locations. Here, Cas9 can be employed to create double-stranded DNA breaks, generate precise mutations, incorporate specific sequences, or regulate RNA transcription. Thus, CRISPR/Cas9-based techniques are of substantial interest for the development of novel therapies to treat genetic diseases. Moreover, as they can be targeted to precise genomic targets with high specificity, they have considerable therapeutic potential for currently “un-druggable” pathologies. Unfortunately, one of the major hurdles in the development of CRISPR/Cas9-based therapies is the intracellular delivery of Cas9-sgRNA complexes. Cas9-sgRNA complexes are unable to cross the cell membrane due to their large size, as well as their negative charge. Moreover, Cas9-sgRNA complexes need to be shielded from opsonization and degradation in circulation, due to their immunogenicity.

To address these problems, your project will focus on the development of extracellular vesicle (EV)-mediated strategies for CRISPR/Cas9 delivery. EVs are lipid membrane nanovesicles released by cells that play an important role in intercellular communication. EVs hold considerable potential for delivery of biotherapeutics, due to their remarkable intrinsic capability of intercellular RNA and protein transfer, as well as their low immunogenicity and their potential for targeted tissue delivery. To harness EVs for CRISPR/Cas9 delivery, you will focus on the development of new strategies that facilitate targeted EV cargo loading and delivery. Techniques that the PhD candidate will employ include EV isolation techniques, CRISPR/Cas-based activity (including genetic engineering and transcriptional activation), the design and cloning of fusion proteins, and cell-based assays and analysis.  

You will take part in the PhD program Drug Innovation of the Graduate School of Life Sciences, which will give you the opportunity to connect with other PhD students in your field and to follow courses.

We are looking for a candidate who meets the following requirements:

• a Master’s degree in Biology, Biomedical Sciences, Pharmaceutical Sciences, or a related field;
• a background in at least one of the following areas: extracellular vesicles, cell biology, cloning/protein engineering, CRISPR/Cas;
• preferable experience with cell culture and analysis (e.g. Western Blot, Microscopy, Flow Cytometry, and/or qPCR);
• excellent communicative skills in English, both in writing and speech.

• the opportunity to work in a collaborative, social, and dedicated team of researchers;
• a position for 4 years;
• a full-time gross salary starting at €2,395 in the first year and increasing to €3,061 in the fourth year (scale P Collective Labour Agreement Dutch Universities (cao));
• benefits including 8% holiday bonus and 8.3% end-of-year bonus;
• a pension scheme, partially paid parental leave, and flexible employment conditions based on the Collective Labour Agreement Dutch Universities.

In addition to the employment conditions laid down in the CAO for Dutch Universities, Utrecht University has a number of its own arrangements. For example, there are agreements on professional development, leave arrangements, and sports. We also give you the opportunity to expand your terms of employment via the Employment Conditions Selection Model. This is how we like to encourage you to continue to grow.

More information about working at the Faculty of Science can be found here.

The Pharmaceutics division is one of five divisions within the Utrecht Institute for Pharmaceutical Sciences and is an internationally renowned top-research group in the field of drug delivery. The core activity of the department is the design and pre-clinical testing of tailor-made drug delivery systems for site-specific, time- and/or rate-controlled delivery of small molecular weight drugs, therapeutic proteins, nucleic acids (including DNA and siRNA), and antigens for therapy and prevention of life-threatening diseases such as cancer, inflammation, and infectious diseases. The Pharmaceutics team consists of scientists from all over the world who like to work in a multidisciplinary environment in which fundamental research and applied sciences are interconnected.

At the Faculty of Science, there are 6 departments to make a fundamental connection with: Biology, Chemistry, Information and Computing Sciences, Mathematics, Pharmaceutical Sciences, and Physics. Each of these is made up of distinct institutes that work together to focus on answering some of humanity’s most pressing problems. More fundamental still are the individual research groups – the building blocks of our ambitious scientific projects.

Utrecht University is a friendly and ambitious university at the heart of an ancient city. We love to welcome new scientists to our city – a thriving cultural hub that is consistently rated as one of the world’s happiest cities. We are renowned for our innovative interdisciplinary research and our emphasis on inspirational research and excellent education. We are equally well-known for our informal atmosphere and the can-do mentality of our people. This lively and inspiring academic environment attracts professors, researchers, and PhD candidates from all over the globe, making both the University and the Faculty of Science a vibrant international and wonderfully diverse community.