A G to A point mutation, converting the 190th amino acid of Rho from aspartic acid (D) to asparagine (N), was introduced using the GalK pop-in/pop-out recombineering method on RP24-288F10 BAC from BACPAC Resource. A loxP-Neo-loxp cassette that was inserted in the intron downstream of the exon carrying the D190N mutation was removed via cre-mediated recombination.