Fig. 1 | Nature Communications

Fig. 1

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

Fig. 1

Nanoblade-mediated genome editing. a Scheme describing the MLV Gag::Cas9 fusion and the Nanoblade production protocol based on the transfection of HEK-293T cells by plasmids coding for Gag-Pol, Gag::Cas9, VSV-G, BaEVRLess, and the sgRNA. b Top panel, immunofluorescence analysis of γ-H2AX (green), RNA polI (red) in U2OS cells 8 h after being transduced with control Nanoblades or with Nanoblades targeting ribosomal DNA genes. Bottom panel, quantification of γ-H2AX and RNA polI colocalization foci in U2OS cells at different times after Nanoblades transduction or after classical DNA transfection methods (n = 3, error bars correspond to standard deviation). c Dose response of Nanoblades. HEK-293T cells were transduced with increasing amounts of Nanoblades targeting human EMX1 (n = 1 displayed). The exact amount of Cas9 used for transduction was measured by dot blot (in gray). Genome editing was assessed by Sanger sequencing and Tide analysis (in red)

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