Sema4c^tm1Matl

A secretory trap cassette was inserted by gene targeting between exons 12 and 13 of the Sema4C gene (exon 1 counted as the exon containing the start codon), which is upstream of the exons encoding the transmembrane (TM) and intracellular domains of Sema4C. The bicistronic mutant transcript encodes two proteins: a fusion of the extracellular domain of the Sema4C protein to the TM and beta-geo domains of the trap vector, and placental alkaline phosphatase (PLAP). Fusion proteins of endogenous proteins with the TM-beta-geo domain of the secretory trap vector, such as the Sema4C-TM-beta-geo fusion, are retained intracellularly and are not present on the cell surface, thereby generating functional null mutations. The absence of protein expression was confirmed by western blot analysis on cerebellar extracts. (J:167973)