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

Reference:

Nat Commun. 2019 Jan 3;10(1):45

Press release (in french):

Les Nanoblades : des navettes pour opérer le génome

Authors:

Mangeot PE1, Risson V2, Fusil F3, Marnef A4, Laurent E3, Blin J3, Mournetas V5, Massouridès E5, Sohier TJM3, Corbin A3, Aubé F6, Teixeira M7, Pinset C5, Schaeffer L2, Legube G4, Cosset FL3, Verhoeyen E3,8, Ohlmann T3, Ricci EP9,10.

Discover the team Eukaryotic and Viral Translation  directed by Théophile Ohlmann

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.

Link to pubmed/full-text

Author information:

Mangeot PE1, Risson V2, Fusil F3, Marnef A4, Laurent E3, Blin J3, Mournetas V5, Massouridès E5, Sohier TJM3, Corbin A3, Aubé F6, Teixeira M7, Pinset C5, Schaeffer L2, Legube G4, Cosset FL3, Verhoeyen E3,8, Ohlmann T3, Ricci EP9,10.

 

  1. CIRI, Centre International de Recherche en Infectiologie Univ Lyon, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon, F-69007, Lyon, France. philippe.mangeot@inserm.fr.
  2. Institut NeuroMyoGène, CNRS 5310, INSERM U121, Université Lyon 1, Faculté de Médecine Lyon Est, Lyon, 69008, France.
  3. CIRI, Centre International de Recherche en Infectiologie Univ Lyon, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon, F-69007, Lyon, France.
  4. LBCMCP, Centre de Biologie Intégrative (CBI), CNRS, Université de Toulouse, UT3, 118 Route de Narbonne, 31062, Toulouse, France.
  5. I-STEM/CECS, Inserm, UMR861 28 rue Henri Desbruères, 91100, Corbeil Essonnes, France.
  6. LBMC, Laboratoire de Biologie et Modélisation de la Cellule Univ Lyon, ENS de Lyon, Université Claude Bernard Lyon 1, CNRS, UMR 5239, INSERM, U1210, Lyon, 69007, France.
  7. SFR BioSciences, Plateau de Biologie Expérimentale de la Souris (AniRA-PBES), Ecole Normale Supérieure de Lyon, Université Lyon1, CNRS UMS3444 INSERM US8, 69007, Lyon, France.
  8. CIRI, Université Côte d’Azur, INSERM, C3M, 06204, Nice, France.
  9. CIRI, Centre International de Recherche en Infectiologie Univ Lyon, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon, F-69007, Lyon, France. emiliano.ricci@ens-lyon.org.
  10. LBMC, Laboratoire de Biologie et Modélisation de la Cellule Univ Lyon, ENS de Lyon, Université Claude Bernard Lyon 1, CNRS, UMR 5239, INSERM, U1210, Lyon, 69007, France. emiliano.ricci@ens-lyon.org.
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