A loxP site was inserted into intron 2 and a loxP site flanked neomycin resistance gene cassette into intron 4. The neo cassette was removed through cre-mediated recombination. Exons 3 and 4 were removed through subsequent cre-mediated recombination, resulting in a knock-out allele. The two exons encode the DHHC-CRD domain. RT-PCR, Western blots and immunohistochemistry confirmed absence of transcription and translation from this allele.