The present disclosure relates to targeted genome editing in eukaryotic cells or organisms. More particularly, the present disclosure provides compositions for inducing targeted disruption of endogenous genes in eukaryotic cells. The composition may comprise a Cas9/guide RNA complex with a Cas9 protein to which a NLS is linked, and a guide RNA having a crRNA and a tracrRNA. The crRNA may comprise i) a portion to be hybridized with a portion of the tracrRNA, and ii) a portion complementary to a target DNA of the endogenous genes. In some embodiments, the Cas/guide RNA complex may be formed in vitro before being introduced into the eukaryotic cell.