Descrizione del progetto
Un nuovo modo per migliorare il trasferimento genico
La terapia genica e l’editing genetico richiedono il trasferimento di cellule negli acidi nucleici. Tuttavia, vi sono varie strozzature ad ostacolare l’efficacia di tale trasferimento. Tra queste ci sono il sequestro endosomiale, la traslocazione nucleare scarsa e, soprattutto, la ritenzione degli acidi nucleici introdotti in gabbie di membrana a seguito del riconoscimento da parte della proteina denominata fattore barriera all’autointegrazione (barrier-to-autointegration factor, BAF). Gli scienziati del progetto IBAF, finanziato dall’UE, affronteranno tale aspetto e aumenteranno la disponibilità e la mobilità citosolica del DNA trasfettato attraverso la soppressione transitoria del BAF. I risultati del progetto faranno progredire la nostra comprensione di questo meccanismo inesplorato di ritenzione del DNA e forniranno stimolatori di trasfezione in grado di potenziare le piattaforme già esistenti di trasferimento dei geni.
Obiettivo
Safe and efficient delivery of nucleic acids to tissues and cells is a shared challenge in the clinical translation of gene therapy and gene editing. At the intracellular level, DNA delivery is hindered by endo/lysosomal sequestration, inefficient transport into the nucleus and a retention mechanism mediated by the barrier-to-autointegration factor (BAF) protein that detects, clusters and locks away intruding double-stranded DNA in membrane cages. The first two of these intracellular obstacles are well-known bottlenecks and are being addressed by many laboratories. However, preliminary in vitro data suggest the detrimental impact of BAF’s mechanism on transgene expression is underestimated. We hypothesize that transiently suppressing BAF or one of its regulating factors will increase the cytosolic availability and mobility of transfected DNA, facilitating its transport to the nucleus and ultimately boosting transfection efficiency. We will tackle this rather uninvestigated mechanism through two parallel strategies: i) identifying novel small-molecule inhibitors of BAF or its regulators via high-throughput screening of chemical libraries, and ii) producing a recombinant kinase that phosphorylates BAF in situ and thus supresses its DNA-clustering function. The BAF suppressors will be co-delivered to cells with rationally-designed nucleoprotein nanoparticles consisting of a reporter plasmid and a dual-function fusion protein to facilitate endocytosis and endosomal escape. The combined transfection-enhancing effect of the BAF suppressors and the nanoparticles will be extensively characterized in vitro, and in vivo proof-of-concept will be obtained in mice. Besides expanding our fundamental knowledge on this unexplored DNA retention mechanism, the project will provide powerful transfection enhancers that can boost already-existing gene delivery platforms (viral and non-viral), so that these can reach their full potential for gene therapy and gene-editing applications.
Campo scientifico
CORDIS classifica i progetti con EuroSciVoc, una tassonomia multilingue dei campi scientifici, attraverso un processo semi-automatico basato su tecniche NLP.
CORDIS classifica i progetti con EuroSciVoc, una tassonomia multilingue dei campi scientifici, attraverso un processo semi-automatico basato su tecniche NLP.
- natural sciencesbiological sciencesbiochemistrybiomoleculesnucleic acids
- medical and health sciencesmedical biotechnologygenetic engineeringgene therapy
- natural sciencesbiological sciencesgeneticsDNA
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- engineering and technologynanotechnologynano-materials
Parole chiave
Programma(i)
Argomento(i)
Meccanismo di finanziamento
ERC-ADG - Advanced GrantIstituzione ospitante
8092 Zuerich
Svizzera