Long Alyssa B, Wilson Isabella M, Terry Tiffany T, Van Sciver Robert E, Caspary Tamara
Department of Human Genetics, Emory University School of Medicine, 615 Michael Street, Suite 301, Atlanta, GA 30322, USA.
Graduate Program in Molecular Biology, Emory University School of Medicine, 615 Michael Street, Suite 301, Atlanta, GA 30322, USA.
J Cell Sci. 2025 Sep 8. doi: 10.1242/jcs.264009.
ARL13B is a regulatory GTPase enriched in cilia, making it a popular marker for this organelle. Arl13bhnn/hnn mice lack ARL13B expression, die during midgestation, and exhibit defects in ciliogenesis. The R26Arl13b-Fucci2aR biosensor mouse line directs the expression of fluorescently tagged full-length Arl13b cDNA upon Cre recombination. To determine whether constitutive, ubiquitous expression of ARL13B-Cerulean can replace endogenous gene expression, we generated Arl13bhnn/hnn animals expressing ARL13B-Cerulean. We show that Arl13bhnn/hnn;Arl13b-Cerulean mice survive to adulthood with no obvious physical or behavioral defects, indicating that the fluorescently tagged protein can functionally replace the endogenous protein during development. However, we observed that rescued males failed to sire offspring, revealing a role for ARL13B in spermatogenesis. This work shows that the R26Arl13b- Fucci2aR mouse contains an inducible allele of Arl13b capable of functioning in most tissues and biological processes.
ARL13B是一种富集于纤毛中的调节性GTP酶，使其成为这种细胞器的常用标志物。Arl13bhnn/hnn小鼠缺乏ARL13B表达，在妊娠中期死亡，并在纤毛发生过程中表现出缺陷。R26Arl13b-Fucci2aR生物传感器小鼠品系在Cre重组后指导荧光标记的全长Arl13b cDNA的表达。为了确定ARL13B-天蓝蛋白的组成型、普遍表达是否可以替代内源性基因表达，我们构建了表达ARL13B-天蓝蛋白的Arl13bhnn/hnn动物。我们发现Arl13bhnn/hnn;Arl13b-天蓝蛋白小鼠存活至成年，没有明显的身体或行为缺陷，这表明荧光标记的蛋白在发育过程中可以在功能上替代内源性蛋白。然而，我们观察到获救的雄性未能产生后代，这揭示了ARL13B在精子发生中的作用。这项工作表明，R26Arl13b-Fucci2aR小鼠包含一个可诱导的Arl13b等位基因，能够在大多数组织和生物学过程中发挥作用。
