title: Highly efficient in vitro and in vivo delivery of functional RNAs using new versatile MS2-chimeric retrovirus-like particles.
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.
authors.affiliation: Université François Rabelais de Tours, INSERM UMR 966 , Tours, France ; UMR UPS/CNRS 5273, EFS-PM, INSERM U1031 , Toulouse, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Prel
authors.affiliation: Université François Rabelais de Tours, INSERM UMR 966 , Tours, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Caval
authors.affiliation: Vectalys, Bâtiment Canal Biotech 2, Parc Technologique du Canal 3 , Toulouse, France.
authors.forename: Régis
authors.initials: R
authors.lastname: Gayon
authors.affiliation: Institut du Cerveau et de la Moelle (ICM), Université Pierre et Marie Curie, CNRS UMR7225; INSERM U1127, Biotechnologies and Biothérapies Team , Paris, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Ravassard
authors.affiliation: Vectalys, Bâtiment Canal Biotech 2, Parc Technologique du Canal 3 , Toulouse, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Duthoit
authors.affiliation: CEA/Université Paris Sud (UMR-E 007), Institut of Emerging Diseases and Innovative Therapies (iMETI), CEA de Fontenay aux Roses , Fontenay aux Roses, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Payen
authors.affiliation: CEA/Université Paris Sud (UMR-E 007), Institut of Emerging Diseases and Innovative Therapies (iMETI), CEA de Fontenay aux Roses , Fontenay aux Roses, France.
authors.forename: Leila
authors.initials: L
authors.lastname: Maouche-Chretien
authors.affiliation: INSERM UMRS 1064, Centre Hospitalier Universitaire (CHU) Hôtel Dieu , Nantes, France ; Institut de Transplantation Urologie Néphrologie (ITUN), Université de Nantes , Nantes, France.
authors.forename: Alison
authors.initials: A
authors.lastname: Creneguy
authors.affiliation: INSERM UMRS 1064, Centre Hospitalier Universitaire (CHU) Hôtel Dieu , Nantes, France ; Institut de Transplantation Urologie Néphrologie (ITUN), Université de Nantes , Nantes, France.
authors.forename: Tuan Huy
authors.initials: TH
authors.lastname: Nguyen
authors.affiliation: Vectalys, Bâtiment Canal Biotech 2, Parc Technologique du Canal 3 , Toulouse, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Martin
authors.affiliation: Université François Rabelais de Tours, INSERM UMR 966 , Tours, France ; CHRU de Tours, Laboratoire de biochimie et biologie moléculaire , Tours, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Piver
authors.affiliation: Vectalys, Bâtiment Canal Biotech 2, Parc Technologique du Canal 3 , Toulouse, France.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Sevrain
authors.affiliation: Vectalys, Bâtiment Canal Biotech 2, Parc Technologique du Canal 3 , Toulouse, France.
authors.forename: Lucille
authors.initials: L
authors.lastname: Lamouroux
authors.affiliation: CEA/Université Paris Sud (UMR-E 007), Institut of Emerging Diseases and Innovative Therapies (iMETI), CEA de Fontenay aux Roses , Fontenay aux Roses, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Leboulch
authors.affiliation: UMR UPS/CNRS 5273, EFS-PM, INSERM U1031 , Toulouse, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Deschaseaux
authors.affiliation: Vectalys, Bâtiment Canal Biotech 2, Parc Technologique du Canal 3 , Toulouse, France.
authors.forename: Pascale
authors.initials: P
authors.lastname: Bouillé
authors.affiliation: UMR UPS/CNRS 5273, EFS-PM, INSERM U1031 , Toulouse, France.
authors.forename: Luc
authors.initials: L
authors.lastname: Sensébé
authors.affiliation: Université François Rabelais de Tours, INSERM UMR 966 , Tours, France ; CHRU de Tours, Laboratoire de biochimie et biologie moléculaire , Tours, France.
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Pagès