HP-F6: 'Oncolytic Viruses' and 'Gene Therapy'
Type: Practical Course with Student Seminars
Weeks 1 and 2: Oncolytic Viruses
Date: 22. January - 2. February 2024
Hosts/Supervisors: Jürg Nüesch (responsible organizer contact: jpf.nuesch@dkfz.de), Mathias Leber, Dirk Nettelbeck, Guy Ungerechts
Topics:
© dkfz.de
Virotherapy represents a novel modality for cancer treatment based on tumor-selective virus infection, replication and spread (therefore also called Viral Oncolysis). Key advantages of this strategy are the implementation of a distinct means of cancer cell killing, amplification of the therapeutic agent in the patients’ tumor, and the opportunity to engineer the virus drug to match specific applications (for example virus capsid modification for targeted cell entry, insertion of therapeutic genes into the viral genome).
Content:
© dkfz.de
Principles of viral oncolysis as well as different oncolytic viruses will be introduced in a morning seminar by the group leaders of DKFZ virotherapy groups. The students will be provided with recent literature, which they will present in the seminar. Practical work will be performed in groups of two students in the different DKFZ virotherapy groups. Students will apply molecular biology, immunology, cell biology and virology techniques to address a scientific topic of oncolytic virus development or characterization.
Week 3: Gene Therapy/Adeno-Associated Virus (AAV)
Date: 10.-14. February 2025
Host: Dirk Grimm and co-workers
Topics and Content:
- Small scale AAV production: detection of capsid proteins via Western Blot, comparison of transduction efficiency of different AAV serotypes
- Medium scale AAV purification: AAV purification via iodixanol gradient ultracentrifugation, infection of different cell lines
- In vitro capsid evolution via DNA family shuffling: DNaseI digest of AAV capsids, shuffling & amplification PCRs, cloning and sequencing of single clones
- RNAi in gene therapy: transfection of different triggers/inhibitors of RNAi, read-out via luciferase reporter assay
- Genome engineering using the CRISPR/Cas9 system: transfection of cells, T7 endonuclease assay