Gatad2b^{tm1a(EUCOMM)Hmgu}

The L1L2_Bact_P cassette was inserted at position 90255407 of Chromosome 3 upstream of the critical exons 2 and 3 (Build GRCm39). The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by a neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of exon 3 at position 90257058. Exons 2and 3 are thus flanked by loxP sites. A "conditional ready" (floxed) allele can be created by flp recombinase expression in mice carrying this allele. Subsequent cre expression results in a knockout mouse. If cre expression occurs without flp expression, a reporter knockout mouse will be created. Further information on targeting strategies used for this and other IKMC alleles can be found at http://www.informatics.jax.org/mgihome/nomen/IKMC_schematics.shtml. RT-qPCR showed that heterozygotes express ~50% of mRNA relative to wild-type controls. Western blot analysis of E18 cerebella showed ~50% of protein content in heterozygotes and absence of protein expression in homozygotes. Immunostaining of forebrain sections from E16.5 embryos confirmed the reduced and absent expression in the nuclei of cells in the cortex of heterozygous and homozygous embryos, respectively. (J:157065, J:344365)