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.

Source: 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 & Emiliano P. Ricci. Nature Communications volume 10, Article number: 45 (2019)