Extended Data Fig. 1: In vitro validation of PrP reduction with CBE-mediated PRNP R37X installation.
From: In vivo base editing extends lifespan of a humanized mouse model of prion disease
a, Representative flow cytometry gating plot showing the fluorescence signal in HEK293T cells treated with BE4max, Cas9 nuclease or dead (deaminase-inactive) BE4max editor, with either PRNP R37X sgRNA (blue) or non-targeting sgRNA (red). Six days after plasmid transfection, cells were incubated with a fluorescently conjugated 6D11 antibody that binds PrP and were analyzed by flow cytometry. b, Mean fluorescence intensity (MFI) of the HEK293T cells after staining with 6D11 antibodies. MFI are plotted after normalizing the MFI of cells treated with PRNP R37X sgRNAs to MFI of cells treated with non-targeting sgRNAs. c, Frequency of the PRNP R37X edit and indels in HEK293T cells after plasmid transfection with BE4max, Cas9 nuclease or dead BE4max, with PRNP R37-targeting sgRNA. Dots represent individual biological replicates (n=3) and data are presented as mean values +/- 95% CI.
