Methods and compositions are provided for nuclease-mediated gene editing of primary cells without the use of viral mediated delivery. Methods of treatments using edited primary cells are also provided.本發明提供在不使用經病毒介導的傳遞之情況下初代細胞的經核酸酶介導之基因編輯之方法及組合物。亦提供使用經編輯初代細胞治療之方法。