Figure 2

Characterisation of X shredder-mCherry and germline promoter-Cas9 transgenic mice. (a) Transgene copy number of hemizygous X-B (n = 2), X-C-1 (n = 3), X-C-2 (n = 3) (two founder lines), and X-D (n = 10) mice. Bars show mean ± SD. (b) Fluorescence imaging performed on ear skin punch biopsies for the transgenic mouse lines described in A. showing representative mCherry signal (red) with a WT mCherry negative control. n =  > 30 per genotype. Scale bar = 200 µM. (c) Expression of Cas9 RNA in testis isolated from Ccna1-, Prm1-, and Stra8-Cas9 transgenic mouse lines. Expression is normalised to eEF2 and WT testis indicates background Cas9 detection in this assay. n = 1 per genotype. (d) Representative IF of Cas9-GFP expression (green) in the testis of Ccna1-Cas9-GFP transgenic mice. GFP signal was amplified by staining with an anti-GFP antibody while DAPI nuclear staining (blue) shows tubule structures. n = 2 per genotype, mice 8–24 weeks of age. Scale bar = 100 µM.