Successful targeting of the <i>Gng5</i> locus.

<p><b>A.</b> Region of mouse chromosome 3∶146,110,000–147,170,000 containing the <i>Gng5</i> locus from the UCSC genome browser. <b>B.</b> The top bar of this schematic illustrates the arrangement of exons of <i>Gng5</i> (orange boxes), <i>Ctbs</i> (blue boxes), and <i>Spata1</i> (yellow boxes). The bars underneath illustrate the <i>Gng5</i> mRNA transcript, the two <i>Ctbs</i> splice variants, the two <i>Ctbs-Gng5</i> splice variants, and the three <i>Spata1</i> splice variants (not drawn to scale). Letters indicate RT-PCR primers which can be found in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0090970#pone.0090970.s003" target="_blank">Table S1</a>. <b>C.</b> RT-PCR of the <i>Ctbs-Gng5</i> fusion transcript (primers <b>f</b> and <b>g</b>) from the same embryos shown in Fig. 1E, confirming that expression of this fusion transcript is preserved in knockout embryos. Identification of the amplified products marked <i>Ctbs-Gng5</i> was confirmed by DNA sequence analysis (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0090970#pone.0090970.s001" target="_blank">Figure S1</a>). The first lane on both gels is a molecular weight marker, φX digested with HaeIII. <b>D.</b> RT-PCR of <i>Spata1</i> from the same embryos (primers <b>f</b> and <b>h</b>), demonstrating that expression of <i>Spata1</i> is preserved in knockout embryos.