Journal: bioRxiv

Article Title: In vivo expansion of gene-targeted hepatocytes through transient inhibition of an essential gene

doi: 10.1101/2023.07.26.550728

Figure Lengend Snippet: a , Eight-week-old C57BL/6J mice were injected with 5×10 11 GC of AAV-CRISPR and 1×10 11 GC of an AAV-Donor containing the promoterless coding sequences of human FAH (selectable marker) and FIX or 6×10 11 GC of AAV-GFP as control. Mice were injected with Fah -siRNA (3 mg/kg or saline as control) every 4 weeks until 12 weeks post-AAV injection. Blood was collected at time 0 and every two to four weeks and mice were sacrificed at 52 weeks post AAV-injection. Created with BioRender.com. b , Human FIX ELISA in plasma samples. c , Representative immunohistochemistry staining of 2A-tagged FAH (FAH-2A), hematoxylin and eosin (H&E) and Fah in the liver. Scale bar is 100 µm. Data are expressed as mean ± s.d. (n= 3 Control and 9 Unselected and Repair Drive mice) with significance determined by two-way ANOVA followed by Tukey test. **** p<0.0001 Repair Drive vs. Control and Unselected mice.

Article Snippet: Human FIX was measured in 1:16.7 diluted plasma by using Factor IX Human SimpleStep ELISA Kit (ab188393, Abcam), following the manufacturer’s protocol.

Techniques: Injection, CRISPR, Marker, Control, Saline, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Immunohistochemistry, Staining