A modified D3 ES cell line heterozygous for the Selptm1Hyn allele was targeted a second time. The exons encoding the signal peptide, lectin domain, and a portion of the epidermal growth factor domain were replaced by the insertion of a PGK-hygro cassette. Both Northern and RT-PCR analysis revealed an absence of normal mRNA for E-selectin but the presence of mRNA for P-selectin in extracts from cardiac and pulmonary tissue of homozygous mutant mice. This targeted mutation occurred in trans with the Selptm1Hyn mutation and were subsequently bred apart. (J:31626)