AbstractWild mouse strains have been used for many research studies, because of the high level of inter-strain genetic and phenotypic variations in them, in addition to the characteristic phenotype maintained from wild mice. However, since application of the current genetic engineering method on wild strains is not easy, there are limited studies that have attempted to apply gene modification techniques in wild strains. Recently, i-GONAD, a new method for genome editing that does not involve any ex vivo manipulation of unfertilized or fertilized eggs has been reported. We applied i-GONAD method for genome editing on a series of wild strains and showed that genome editing is efficiently possible using this method. We successfully made genetically engineered mice in seven out of the nine wild strains. Moreover, we believe that it is still possible to apply milder conditions and improve the efficiencies for the remaining two strains. These results will open avenues for studying the genetic basis of various phenotypes that are characteristic to wild strains. Furthermore, applying i-GONAD will be also useful for other mouse resources in which genetic manipulation is difficult using the method of microinjection into fertilized eggs.