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Vous êtes ici : Accueil / Équipes / Métabolisme des ARNs au cours de la réponse immunitaire et de l’infection - E. Ricci / Publications / Genome editing in primary cells and in vivo using viral-derived Nanoblades loaded with Cas9-sgRNA ribonucleoproteins.

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

Philippe E Mangeot, Valerie Risson, Floriane Fusil, Aline Marnef, Emilie Laurent, Juliana Blin, Virginie Mournetas, Emmanuelle Massourides, Thibault JM Sohier, Antoine Corbin, Fabien Aube, Marie Teixeira, Christian Pinset, Laurent Schaeffer, Gaelle Legube, Francois-Loic Cosset, Els Verhoeyen, Theophile Ohlmann, and Emiliano P Ricci (2019)

Nat Commun, 10(1):45.

Programmable nucleases have enabled rapid and accessible genome engineering in eukaryotic cells and living organisms. However, their delivery into target cellscan 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 linesand 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 issimple, relatively inexpensive and can be easily implemented in any laboratory equipped for cellular biology.

 
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