Genome editing in primary cells and in vivo using viral-derived Nanoblades loaded with Cas9-sgRNA ribonucleoproteins
Résumé
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.
Domaines
Biologie moléculaire| Origine | Fichiers éditeurs autorisés sur une archive ouverte |
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