Fig. 6: Systemic injection of Prpf31-KO vectors to P0 mice. | Nature Communications

Fig. 6: Systemic injection of Prpf31-KO vectors to P0 mice.

From: Gene augmentation prevents retinal degeneration in a CRISPR/Cas9-based mouse model of PRPF31 retinitis pigmentosa

Fig. 6

a Illustration of the systemic injection approach. AAV92YF-CMV-SaCas9-U6-gRNAg/h (50 µl, 1.35E + 13 vg/ml, gRNAg:gRNAh = 1:1) was delivered to P0 WT mice via facial vein injection. An equal volume of PBS or mock Cas9 vector AAV92YF-CMV-SaCas9-Nt gRNA (3.82E + 13 vg/ml) was injected to litter mates as control. b Images of 2 male mice taken 4 weeks (p.i.) Compared with PBS-injected control litter mate, KO pups developed abnormally. c Body weights of PBS, Mock and KO vectors injected and uninjected pups 2–4 weeks (p.i.) The KO group had significantly lower body weights (n = 3–6). Males and females were evenly distributed in each group. (2 weeks: PBS n = 5, Mock n = 4, KO n = 6; 3 weeks: PBS n = 4, Mock n = 4, KO n = 3; 4 weeks: PBS n = 4, Mock n = 4, KO n = 5). d Survival plot of PBS, Mock and KO-injected pups. KO pups had a high early mortality rate (PBS n = 16, Mock n = 4, KO n = 18). The first check point was at P14 to avoid disturbing dams and pups. e PCR amplification of gRNA targeted region showed truncated Prpf31 (black arrow) in DNA collected from the KO group. f, g Western blots of liver samples collected from PBS, Mock and KO vectors injected pups. Bar plots of relative PRPF31 band intensity normalized by GAPDH show a significant reduction of PRPF31 protein in the KO group (n = 3). Data were analyzed by unpaired two-tailed t-test and are shown as mean ± SEM. *P < .05; **P < .01. Source data are provided as a Source Data file.

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