A targeting vector was designed to insert a stop codon into exon 6 at amino acid 121, as well as replace exons 6-8 of the targeted gene with an internal ribosome entry site-b-galactosidase (IRES-lacZ) fusion gene and neomycin cassette. The construct was electroporated into 129SvEv-derived embryonic stem (ES) cells. RT-PCR analysis of brain tissue in homozygotes confirms lack of normal mRNA expression, while X-gal staining closely mimics dynamic expression pattern of normal Nkd2 mRNA. (J:122353)