Fig. 5: Engineering tissue-specific expression of dual-AAV TadCBEd.
From: In vivo base editing extends lifespan of a humanized mouse model of prion disease
a,b, Frequency of the desired R37X edit (a), and PrP protein level in the bulk brain hemisphere (b) of Tg25109 mice treated with dual-AAV PHP.eB TadCBEd PRNP R37X F+E-sgRNA, with Cbh (n = 12), hSYN (n = 11) or EFS (n = 6) promoter driving the expression of the base editor. Data for the ‘Cbh’ condition correspond to the ‘TadCBEd+PRNP R37X F+E-sgRNA’ condition in Fig. 3d,e, and are replotted for comparison. c, Frequency of the desired R37X edit in the liver of mice untreated (n = 6) or treated with dual-AAV TadCBEd encoding Cbh promoter (Cbh; n = 6), hSYN promoter (hSYN; n = 6), hSYN promoter plus miR-183 target sites (hSYN+miR-183; n = 6), hSYN promoter plus miR-122 target sites (hSYN+miR-122; n = 5) or hSYN promoter plus miR-183 and miR-122 target sites (hSYN+miR-183+miR-122; n = 5). d, Schematic of dual-AAV PHP.eB TadCBEd PRNP R37X F+E-sgRNA with hSYN promoter driving the expression of the base editor and miR target site (TS) incorporation. ‘miR-183’ contains 4 copies of miR-183 target sites; ‘miR-122’ contains 3 copies of miR-122 target sites; ‘miR-183+miR-122’ contains 3 copies each of miR-183 and miR-122 target sites. e,f, Frequency of the desired R37X edit (e) (P < 0.0001 for all groups versus untreated), and PrP protein level (f) in the bulk brain hemisphere of Tg25109 mice harvested 35 d after treatment with dual-AAV TadCBEd, with or without the specified miR target site incorporation (untreated versus hSYN, P = 0.0003; untreated versus hSYN+miR-183, P = 0.0001; hSYN+miR-122, P = 0.0002; untreated versus hSYN+miR-183+miR-122, P < 0.0001) at a total dose of 1.5 × 1013 vg kg−1 (untreated, n = 6; hSYN, n = 11; hSYN+miR-183, n = 6; hSYN+miR-122, n = 5; hSYN+miR-183+miR-122, n = 5). Data for the ‘hSYN’ condition correspond to the ‘hSYN’ condition in a and b, and are replotted for comparison. Dots represent individual biological replicates and data are presented as mean ± 95% CI. Significance in a and b was calculated by one-way ANOVA test with Bonferroni correction; NS, P > 0.12. Significance in e and f was calculated by two-way ANOVA test with Dunnett’s correction; ***P < 0.0002; ****P < 0.0001.
