逆转录病毒
体外
体内
细胞生物学
计算生物学
化学
病毒学
生物
病毒
遗传学
作者
Anne Prel,Vincent Caval,Régis Gayon,Philippe Ravassard,Christine Duthoit,Emmanuel Payen,Stany Chrétien,Alison Crénéguy,Tuan Huy Nguyen,Nicolas Martin,Éric Piver,Raphaël Sevrain,Lucille Lamouroux,Philippe Leboulch,Frédéric Deschaseaux,Pascale Bouillé,Luc Sensebé,Jean‐Christophe Pagès
摘要
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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