Figure 1.

In Vivo Genome Editing of Albumin Locus in Mouse Liver by AAV.CRISPR-Cas9

(A) Schematic illustrating albumin targeting strategy. (B) In vitro validation of sgRNAs targeted to mAlb1 in the H2.35 mouse cell line by transient transfection and Surveyor nuclease assays. sgRNA4 showed the highest efficiency in inducing indels in the targeted loci and was therefore chosen for subsequent studies. Arrows denote Surveyor nuclease cleaved fragments of the Alb PCR products. Results were replicated in two independent experiments. (C) Dual AAV vector system for liver-directed and SpCas9-mediated gene insertion. The AAV8.sgRNA.donor vector encoded a promoterless hFIX cassette containing exons 2–8 of the hFIX gene flanked by a SA signal and a poly(A) (PA) sequence.