PROJECT INFORMATION
Host institution: Aarhus University, Aarhus
Country: Denmark
Supervisory team: Prof. Jacob Giehm Mikkelsen (Aarhus University, Denmark) Prof. Peter Loskill (Universität Tubingen, Germany), Assistant Prof Marianne Carlon (KU Leuven, Belgium)
PROJECT DESCRIPTON
The objectives are:
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To establish LVNP-directed gene modification by CRISPR/Cas9 and CRISPRi in lung cell and tissue models.
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To achieve targeted genome modification using LVNPs based on panel of different pseudotypes in different model systems.
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To identify genes influencing intracellular transport and protective cellular responses during LVNP-directed gene editor delivery in lung cell models.
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To modulate potential innate immune signalling pathways and DNA damage responses to optimize LVNP delivery in lung tissue.
In this project, we will develop lentivirus-derived nanoparticles (LVNPs) for CRISPR/Cas-based gene editing in lung tissue, targeting innate immune signaling and DNA damage responses.
LVNPs will be compared to other VLP technologies and pseudotyping will be studied for targeting of ACE2+ cells. Delivery efficacy will be tested in ex vivo EVLP and in vitro ischemia-reperfusion injury lung models. Cellular responses to LVNPs, including innate immune activation and DNA damage responses, will be characterized. Genome-wide CRISPR screens will identify genes that modulate delivery efficiency, informing optimal strategies for lung gene therapy.
A successful project will result in: (a) Establishment of LVNP technology for robust targeted gene knockout and knockdown in lung cell models and lung models. (b) Identification of cellular factors that either support or restrict LVNP-directed gene editing in lung tissue leading to optimized delivery of gene editing tool kits to lung tissue.
Enrolment in Doctoral School: The Graduate School of Health, Faculty of Health, Aarhus University
Planned secondments:
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Miltenyi Biotech, Germany: MACSima system for multiplex imaging of LVNP-treated cells (month 10).
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Universität Tubingen, Germany: Organ-on-a-chip model for studies of LVNP delivery (months 14-5)
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KU Leuven, Belgium: LVNP-directed CRISPR/Cas RNP delivery in (precision cut lung slices and EVLP models (months 23-25)
ESSENTIAL REQUIREMENTS
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You hold a Master’s degree (no PhD) in molecular biology, biomedical sciences, biotechnology, or similar.
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You are passionate about life sciences and engineering, and want to achieve a PhD degree on the topic described in the description above
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You are ambitious, well organized, and have excellent communication skills
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You are proficient in English both spoken and written
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You have a solutions-oriented mindset that thrives in a multidisciplinary team
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You can work independently and have a critical mindset.
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You are an enthusiastic and motivated person, eager to participate in network-wide training events, international travel and public awareness activities.
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You are willing to travel and eager to interact with other PhD students across Europe.
DESIRABLE SKILLS AND EXPERTISE
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You are experienced in the use of CRISPR/Cas-based gene editing technologies
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You have solid expertise in virus-based gene and protein delivery technologies, in particular retro- or lentiviral systems
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You have hands-on experience with diverse molecular biology techniques