Combinatorial testing of viral vector and CRISPR systems for precision genome editing

Li, Z. (2025)

Doctoral Thesis

The work present in this dissertation provides the evidence that delivery of well-established CRISPR/Cas9 gene editing machinery rationally allocated in different viral vectors can achieve plausibly ideal results of gene editing in the mammalian cells regarding efficiencies, accuracy and specificity. The use of CRISPR/Cas9 system makes the gene editing convenient and customized and compared to other delivery options, the viral vectors are outperformed due to natural evolution coffering high compatibility to mammalian cells. However, it would still be a long road to go along with all kinds of obstacles, then finally we may test in one clinical trial the concept of such combination investigated in the thesis.

• Precision gene editing
• CRISPR/Cas9
• High-capacity adenoviral vector
• Adeno-associated viral vector
• Homology-directed repair

• All authors: Li, Z.
• Supervisor: Hoeben, R.C.
• Co-supervisor: Gonçalves, M.A.F.V.
• Committee: Chuva de Sousa Lopes, S.M.; Pijnappel, W.W.M.; Heemskerk, M.H.M.; Dijke, P. ten; Lei, Z.Y.
• Qualification: Doctor (dr.)
• Awarding Institution: Faculty of Medicine, Leiden University Medical Center (LUMC), Leiden University
• Date: 2025-10-08
• ISBN (print): 9789465225685
• Sponsorship: the China Scholarship Council (CSC)-Leiden University joint program; the Prinses Beatrix Spierfonds; the Duchenne Parent Project NL; the Dutch Research Council (NWO)-OpenTechnology Program; the EU Marie Skłodowska-Curie Doctoral Network Actions