Genome editing in primary cells and in vivo using viral-derived Nanoblades loaded with Cas9-sgRNA ribonucleoproteins

Philippe E. Mangeot (), Valérie Risson, Floriane Fusil, Aline Marnef, Emilie Laurent, Juliana Blin, Virginie Mournetas, Emmanuelle Massouridès, Thibault J. M. Sohier, Antoine Corbin, Fabien Aubé, Marie Teixeira, Christian Pinset, Laurent Schaeffer, Gaëlle Legube, François-Loïc Cosset, Els Verhoeyen, Théophile Ohlmann and Emiliano P. Ricci ()
Additional contact information
Philippe E. Mangeot: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon
Valérie Risson: Faculté de Médecine Lyon Est
Floriane Fusil: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon
Aline Marnef: Université de Toulouse
Emilie Laurent: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon
Juliana Blin: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon
Virginie Mournetas: Inserm
Emmanuelle Massouridès: Inserm
Thibault J. M. Sohier: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon
Antoine Corbin: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon
Fabien Aubé: Université Claude Bernard Lyon 1, CNRS, UMR 5239, INSERM, U1210
Marie Teixeira: Université Lyon1, CNRS UMS3444 INSERM US8
Christian Pinset: Inserm
Laurent Schaeffer: Faculté de Médecine Lyon Est
Gaëlle Legube: Université de Toulouse
François-Loïc Cosset: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon
Els Verhoeyen: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon
Théophile Ohlmann: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon
Emiliano P. Ricci: Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon

Nature Communications, 2019, vol. 10, issue 1, 1-15

Abstract: Abstract Programmable nucleases have enabled rapid and accessible genome engineering in eukaryotic cells and living organisms. However, their delivery into target cells can be technically challenging when working with primary cells or in vivo. Here, we use engineered murine leukemia virus-like particles loaded with Cas9-sgRNA ribonucleoproteins (Nanoblades) to induce efficient genome-editing in cell lines and primary cells including human induced pluripotent stem cells, human hematopoietic stem cells and mouse bone-marrow cells. Transgene-free Nanoblades are also capable of in vivo genome-editing in mouse embryos and in the liver of injected mice. Nanoblades can be complexed with donor DNA for “all-in-one” homology-directed repair or programmed with modified Cas9 variants to mediate transcriptional up-regulation of target genes. Nanoblades preparation process is simple, relatively inexpensive and can be easily implemented in any laboratory equipped for cellular biology.

Date: 2019
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DOI: 10.1038/s41467-018-07845-z

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