Abstract
Recent developments in the precise gene ediঞng field enabled the correcঞon of many different geneঞc diseases in vitro and in vivo. For therapeuঞc applicaঞon, safe and efficient delivery methods of these tools are required. Virus-like parঞcles (VLPs) are DNA-free vesicles taking advantage of viral delivery proteins while bypassing the risk of genomic integraঞon and long-term expression. Here, we produce and opঞmize engineered virus-like parঞcles (eVLPs) as a tool for in vitro delivery of base and prime ediঞng machinery in the form of ribonucleoproteins (RNPs). We showed highly efficient base ediঞng in HEK293T cells and in intrahepaঞc cholangiocyte organoids (ICOs) a[er delivery with eVLPs. Furthermore, we adapted the system for the delivery of prime editors and showed ediঞng in HEK293T cells. Further opঞmizaঞons of the PE-eVLP system allowed us to increase the ediঞng efficiencies with prime ediঞng by 16%. Finally, cysঞc fibrosis colon organoids were chosen to show funcঞonal correcঞon a[er delivery of ediঞng tools, which did not result in ediঞng of the targeted mutaঞons. Altogether, these results support eVLPs as an efficient in vitro delivery system for precise gene ediঞng machinery.
| Original language | English |
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| Publication status | Published - 2023 |
| Event | European Society of Gene and Cell Therapy - Belgium, Brussels, Belgium Duration: 23 Oct 2023 → 27 Oct 2023 |
Conference
| Conference | European Society of Gene and Cell Therapy |
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| Abbreviated title | ESGCT |
| Country/Territory | Belgium |
| City | Brussels |
| Period | 23/10/23 → 27/10/23 |