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Mol Ther Methods Clin Dev. 2015 Oct 21;2:15039. doi: 10.1038/mtm.2015.39. eCollection 2015.

Highly efficient in vitro and in vivo delivery of functional RNAs using new versatile MS2-chimeric retrovirus-like particles.

Author information

1
Université François Rabelais de Tours, INSERM UMR 966 , Tours, France ; UMR UPS/CNRS 5273, EFS-PM, INSERM U1031 , Toulouse, France.
2
Université François Rabelais de Tours, INSERM UMR 966 , Tours, France.
3
Vectalys, Bâtiment Canal Biotech 2, Parc Technologique du Canal 3 , Toulouse, France.
4
Institut du Cerveau et de la Moelle (ICM), Université Pierre et Marie Curie, CNRS UMR7225; INSERM U1127, Biotechnologies and Biothérapies Team , Paris, France.
5
CEA/Université Paris Sud (UMR-E 007), Institut of Emerging Diseases and Innovative Therapies (iMETI), CEA de Fontenay aux Roses , Fontenay aux Roses, France.
6
INSERM UMRS 1064, Centre Hospitalier Universitaire (CHU) Hôtel Dieu , Nantes, France ; Institut de Transplantation Urologie Néphrologie (ITUN), Université de Nantes , Nantes, France.
7
Université François Rabelais de Tours, INSERM UMR 966 , Tours, France ; CHRU de Tours, Laboratoire de biochimie et biologie moléculaire , Tours, France.
8
UMR UPS/CNRS 5273, EFS-PM, INSERM U1031 , Toulouse, France.

Abstract

RNA delivery is an attractive strategy to achieve transient gene expression in research projects and in cell- or gene-based therapies. Despite significant efforts investigating vector-directed RNA transfer, there is still a requirement for better efficiency of delivery to primary cells and in vivo. Retroviral platforms drive RNA delivery, yet retrovirus RNA-packaging constraints limit gene transfer to two genome-molecules per viral particle. To improve retroviral transfer, we designed a dimerization-independent MS2-driven RNA packaging system using MS2-Coat-retrovirus chimeras. The engineered chimeric particles promoted effective packaging of several types of RNAs and enabled efficient transfer of biologically active RNAs in various cell types, including human CD34(+) and iPS cells. Systemic injection of high-titer particles led to gene expression in mouse liver and transferring Cre-recombinase mRNA in muscle permitted widespread editing at the ROSA26 locus. We could further show that the VLPs were able to activate an osteoblast differentiation pathway by delivering RUNX2- or DLX5-mRNA into primary human bone-marrow mesenchymal-stem cells. Thus, the novel chimeric MS2-lentiviral particles are a versatile tool for a wide range of applications including cellular-programming or genome-editing.

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