Fig. 3: In vivo Adenine Base Editing upon low dose AAV split-vector administration.

A Depiction of the split-vector system. AAV_SV1 contains a U6 promoter-driven gRNA gene and the LP1-driven N-terminal ABE7.10 domain. AAV_SV2 contains the C-terminal part. B Experimental timeline. Basal blood was withdrawn on day 0. Split-vector was injected on day 1. On day 14 animals were supplied with iron-enriched food until the endpoint. A second blood withdrawal occurred on day 28 and a third on day 112. Editing efficiency was assessed by NGS and iron accumulation was determined via Prussian Blue staining. C Determination of base conversion rate via NGS four months after low-dose AAV8 split-vector injection. The bars in the scatter dot plot depict the mean with the SD. Male mice are represented in circles, female mice in triangles. Hfe locus was PCR-amplified from liver DNA prior to NGS (n = 5 controls, n = 8 treated). D Transferrin saturation was measured from serum withdrawn at day 28 and 112. Statistical significance was determined using a two-tailed unpaired t test. p value = 0.0106 (AAV8 Split vector day 28 vs AAV8 Split vector day 112). Data are presented as mean values +/− SEM. TS% Day 28 85.80 ± 3.731, n = 6; TS% Day 112 70.21 ± 3.290, n = 6. The bars in the scatter dot plot depict the mean with the SD. Male mice are represented in circles, female mice in triangles. E UIBC was measured from serum obtained at day 28 and 112. Statistical significance was determined using a two-tailed unpaired t test. p value = 0.0147 (AAV8 Split vector day 28 vs AAV8 Split vector day 112). Data are presented as mean values +/− SEM. UIBC Day 28: 8.700 ± 2.852; n = 6; UIBC Day 112: 22.84 ± 3.798, n = 7. The bars in the scatter dot plot depict the mean with the SD. Male mice are represented in circles, female mice in triangles. F Hepcidin levels were measured from serum obtained at the endpoint. Statistical significance was determined using a two-tailed unpaired t test. p value = 0.0199. Data are presented as mean values +/− SEM. AAV8 Split vector: 451.7 ± 54.06, n = 10; Control: 254.2 ± 33.45, n = 6. The bars in the scatter dot plot depict the mean with the SD. Male mice are represented in circles, female mice in triangles. G H&E staining of liver sections and hepatic iron accumulation four months post low-dose split-vector injection. Liver sections were stained with Prussian Blue and densitometric quantification was performed Statistical significance was determined using a two-tailed unpaired t test. p value = 0.0251. Data are presented as mean values +/− SEM. AAV8 Split vector: 0.1909 ± 0.02375, n = 13; Control: 0.4191 ± 0.09250, n = 13. The bars in the scatter dot plot depict the mean with the SD. Male mice are represented in circles, female mice in triangles. Source data are provided as a Source Data file.