Journal
SCIENTIFIC REPORTS
Volume 8, Issue -, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41598-017-18826-5
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Funding
- SFR Biosciences [US8 INSERM, UMS3444CNRS]
- European Research Council [ERC-2013-CoG_616986]
- Agence Nationale de la Recherche [ANR-13-JSV3-0002-01]
- Agence Nationale de la Recherche Agence Nationale de la Recherche [ANR-12-RPIB-0011]
- SFR Biosciences (UCBL)
- Ligue Nationale Contre leCancer Comite du Rhone
- Association pour la Recherche sur le Cancer [SFI20111204053]
- Ligue Contre leCancer Comites de l'Isere et de Savoie
- SFR Biosciences (ENS de Lyon)
- Agence Nationale de la Recherche (Thyromut2 program) [ANR-15-CE14-0011-01]
- Agence Nationale de la Recherche (ANR) [ANR-12-RPIB-0011, ANR-13-JSV3-0002, ANR-15-CE14-0011] Funding Source: Agence Nationale de la Recherche (ANR)
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In this report, we present an improved protocol for CRISPR/Cas9 genome editing in mice. The procedure consists in the electroporation of intact mouse zygotes with ribonucleoprotein complexes prepared in vitro from recombinant Cas9 nuclease and synthetic dual guide RNA. This simple cloning-free method proves to be extremely efficient for the generation of indels and small deletions by non-homologous end joining, and for the generation of specific point mutations by homology-directed repair. The procedure, which avoids DNA construction, in vitro transcription and oocyte microinjection, greatly simplifies genome editing in mice.
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