Descrizione del progetto
Un nuovo virus sintetico per il trasferimento genico mirato
La capacità dei virus di infettare e penetrare le cellule, e di riprodursi all’interno di esse, ha richiamato un grande interesse nel settore della terapia genica. Il progetto GENESHUTTLE, finanziato dall’UE, mira a creare strutture sintetiche simili a virus che portano informazioni genetiche di interesse e ricapitolano le proprietà dei virus naturali. I ricercatori impiegheranno il DNA a origami per assicurare il corretto ripiegamento dell’acido nucleico, mentre le particelle generate saranno in grado di interagire con le membrane bersaglio e innescare l’internalizzazione evitando tuttavia la replicazione autonoma incontrollata. Il sistema di trasferimento GENESHUTTLE rappresenterà un progresso notevole rispetto agli approcci attualmente disponibili e si prevede che troverà applicazioni immediate nella ricerca e nella medicina.
Obiettivo
Using rational design with DNA origami, we propose to create synthetic virus-like assemblies capable of accomplishing cell-invading and gene expression functionalities known so far only from natural viruses. We envision these assemblies to be useful for studying and testing viral import mechanisms, and also for gene delivery, enabling fundamental studies and potential medical applications. The objectives include selecting and invading a target cell type, systematically solving the challenges of endosomal escape and nuclear delivery, and inducing the expression of user-defined genetic information in the nucleus. To achieve these objectives, we will recreate and experimentally test mechanisms believed to be used by viruses, including receptor-mediated endocytosis, stimulus-dependent lipid membrane penetration, membrane fusion, active cytosolic transport, and nuclear import. We foresee building synthetic shells carrying genetic information stored in nucleic acids, which we refer to as the “gene shuttle”. The particles will optionally include a membrane envelope and user-defined surface features to mediate receptor-ligand interactions with cell membranes, and they will be capable of stimulus-dependent conformational changes to trigger membrane fusion or membrane penetration. They will shed structural elements on the path to the nucleus, similar to viruses. In addition to delivering genes for fluorescent markers, as a proof of concept demonstration we plan to use the gene shuttle to deliver the genetic information for expressing chimeric antigen receptors (CAR) in a T cell line, which promises to be of use in cancer immunotherapy. The project promises to yield a gene delivery system with capabilities beyond current synthetic vectors, which struggle to overcome the many cellular barriers to deliver and express genetic cargo. For safety reasons, the gene shuttle will by design be unable to assemble in the context of a cell to prevent uncontrolled autonomous replication.
Campo scientifico
- natural sciencesbiological sciencesbiochemistrybiomoleculesnucleic acids
- natural sciencesbiological sciencesmicrobiologyvirology
- natural sciencesbiological sciencesbiochemistrybiomoleculeslipids
- medical and health sciencesclinical medicineoncology
- medical and health sciencesbasic medicineimmunologyimmunotherapy
Parole chiave
Programma(i)
Argomento(i)
Meccanismo di finanziamento
ERC-ADG - Advanced GrantIstituzione ospitante
80333 Muenchen
Germania