Figure 1. Targeted disruption of the Taok2 gene in mice.

(a) Schematic of the conditionally targeted Taok2 allele. The Taok2tm1 allele was generated by introduction of lox P sites flanking exons 2 though 7 of the Taok2 gene. Within intron 7 is a neomycin resistance gene flanked by lox P and frt sites. Upon crossing this line to mice expressing Flpe recombinase, the neomycin selectable marker is removed along with one of the lox P sites to generate the Taok2tm1fl allele. In the presence of Cre recombinase, the remaining lox P sites are recombined into a single lox P site, removing exons 2 though 7 and the translational start site to generate the Taok2tm1Δ allele. (b) Brain sagittal section of a control mouse shows strong expression of TAOK2 by immunohistological analysis in all areas, especially in cortex, hippocampus and striatum. (c) Immunohistological analysis shows reduced TAOK2 expression in Taok2tm1fl/fl;Nes-cre brain. (d) Quantitative PCR analysis of total mouse brain extracts shows near absence of Taok2 transcript in Taok2tm1fl/fl;Nes-cre brain (n = 4) compared to control mice (n = 2). (e) Western blot showing comparative loss of TAOK2 protein in total brain lysates of Taok2tm1fl/fl;Nes-cre mice relative to controls. (f) Quantification of anti-TAOK2 signal from (e) comparing Taok2tm1fl/fl;Nes-cre mice (n = 2) and controls (n = 2). Lox P sites are denoted by grey chevrons, frt sites by black chevrons. Neo = neomycin resistance gene. The positions of restriction sites BamH1 and BsrB1 used in the generation of the targeting construct are indicated. CTX = cortex; HC = hippocampus; STR = striatum. Error bars are mean ± SEM. Asterisks indicate level of significance (* P < 0.05).