Fig. 4
In vivo gene editing after oviduct injection with rAAV vectors. a Schematic representation of the strategy to induce in vivo gene editing of the Tyr locus. rAAV vectors carrying SpCas9 and sgTyr expression constructs were injected directly into the oviduct of plugged C57BL/6NJ females mated to C57BL/6NJ males. Coat color was assessed in the offspring. b Close-up view of the reproductive tract showing the process of in vivo delivery of rAAVs into the ampulla of the oviduct. The rAAV injection solution contains a blue tracer dye that is visible at the tip of the glass micropipette. A small pool of the injected solution is evident inside the ampulla. U uterus, Ov oviduct, O ovary. c Representative litter born after in vivo gene editing of Tyr. One out of eight pups born was albino. d Stacked histogram showing the percentage distribution of indel-type frequencies in two albino (1 and 2) and two black (3 and 4) pups by SMRT sequencing. Three C57BL/6NJ control samples (C1–C3) were included in the analysis. Alterations indicate base replacements; compound mutations are combinations of insertions, deletions, and/or alterations. The yellow area in sample C3 is likely the product of sequencing error. e Litter derived from Tyr-edited albino male crossed with a CD-1 female. All pups are albino indicating germline transmission
