Skip to main content
. 2019 Jun 17;116(27):13288–13292. doi: 10.1073/pnas.1903230116

Fig. 1.

Fig. 1.

Generation of targeted gene knockout quail using adenovirus. (A) Schematic representation of research plan. gRNA of the MLPH gene was selected from exon 2, and the adenoviral CRISPR/Cas9 vector was constructed. Recombinant adenovirus was subsequently produced and injected into the quail blastoderm. Chimeras (G0) were maintained and mated with wild-type (WT) quail to produce G1 quail with a heterozygous genotype of the MLPH gene (MLPH+/−). Male and female G1 MLPH+/− quail were subsequently mated to generate G2 quail with a homozygous genotype of the MLPH gene (MLPH−/−). After production of the recombinant adenovirus, it took a total of 10 wk, 2 wk for egg incubation after virus injection and another 8 wk for sexual maturation, to receive eggs from G0 quail. After G1 quail hatched, the MLPH+/− quail were screened and mated to produce G2 offspring 8 wk later. Fully grown G2 offspring were obtained for further experiments 26 wk after the injection of the adenovirus. (B) Sanger sequencing chromatograms of G1 MLPH+/− quail. Dashed lines indicate the starting point of the mutation, and the deleted or inserted nucleotides are underlined. The PAM sequences are highlighted in gray. Nucleotide sequences are presented in a negative direction.