Fig. 4

Creating large deletion using the i-GONAD method. a Schematic diagram showing deletion of 16.2-kb sequence consisting of retrotransposon in the C57BL/6JJcl mouse genome, to restore agouti phenotype. The target sequences and genotyping primers are shown. ssODN containing EcoRI site at the middle of the sequence was used. b Representative mice showing rescued agouti phenotype (indicated by yellow arrows). These mice were recovered through caesarean section and nursed by Jcl:MCH(ICR) foster mother with her own pups. c Genotyping analyses. Expected fragment sizes: M943/M948 = 290 or 295 bp (ssODN knock-in), M943/M944 = 337 bp, M947/M948 = 477 bp. d Direct sequencing results of PCR products amplified from the G0 mice. The position of junctional sequences is indicated by yellow rectangles. e EcoRI digestion of PCR products amplified from G0 mice (G0-#3 and -#5) with the M943/M948 primer set. Red arrow indicates digested fragment. f Efficiencies of agouti gene editing (see Additional file 1: Table S4 for details)