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Updated: Nov 1, 2025
Functional Cloning Using a Xenopus Oocyte Expression System
Published on: January 30, 2016
The highly conserved FOXJ1 target CFAP161 is dispensable for motile ciliary function in mouse and Xenopus
Anja Beckers1, Franziska Fuhl2, Tim Ott2
1Institute for Molecular Biology, OE5250, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany.
Abstract:
Cilia are protrusions of the cell surface and composed of hundreds of proteins many of which are evolutionary and functionally well conserved. In cells assembling motile cilia the expression of numerous ciliary components is under the control of the transcription factor FOXJ1. Here, we analyse the evolutionary conserved FOXJ1 target CFAP161 in Xenopus and mouse. In both species Cfap161 expression correlates with the presence of motile cilia and depends on FOXJ1. Tagged CFAP161 localises to the basal bodies of multiciliated cells of the Xenopus larval epidermis, and in mice CFAP161 protein localises to the axoneme. Surprisingly, disruption of the Cfap161 gene in both species did not lead to motile cilia-related phenotypes, which contrasts with the conserved expression in cells carrying motile cilia and high sequence conservation. In mice mutation of Cfap161 stabilised the mutant mRNA making genetic compensation triggered by mRNA decay unlikely. However, genes related to microtubules and cilia, microtubule motor activity and inner dyneins were dysregulated, which might buffer the Cfap161 mutation.
Insights
The transcription factor FOXJ1 controls motile cilia assembly. We studied CFAP161, a conserved FOXJ1 target, in Xenopus and mice, finding its disruption surprisingly caused no motile cilia defects, suggesting compensatory mechanisms.
Area of Science:
- Cell Biology
- Developmental Biology
- Genetics
Background:
- Cilia are crucial cell protrusions involved in motility and signaling.
- The transcription factor FOXJ1 regulates the expression of many genes essential for motile cilia assembly.
- CFAP161 is an evolutionarily conserved FOXJ1 target gene identified in motile cilia.
Purpose of the Study:
- To investigate the evolutionary conservation and functional role of CFAP161 in motile cilia formation.
- To determine if CFAP161 is essential for motile cilia function in Xenopus and mouse models.
- To explore potential compensatory mechanisms in the absence of CFAP161.
Main Methods:
- Analysis of Cfap161 expression in Xenopus and mouse models.
- Localization studies of tagged CFAP161 protein in multiciliated cells.
- Gene disruption (knockout) of Cfap161 in both species.
- Assessment of motile cilia phenotypes.
- Transcriptomic analysis to identify dysregulated genes.
Main Results:
- Cfap161 expression is conserved and FOXJ1-dependent in motile ciliated cells.
- CFAP161 localizes to basal bodies (Xenopus) and axonemes (mouse).
- Disruption of Cfap161 did not result in observable motile cilia phenotypes in either species.
- Genetic compensation via mRNA decay was unlikely due to mRNA stabilization.
- Genes related to microtubules, cilia, motor activity, and inner dyneins were dysregulated.
Conclusions:
- CFAP161 is a conserved component of motile cilia, regulated by FOXJ1.
- Despite conserved expression and localization, CFAP161 is not essential for motile cilia function in Xenopus and mouse.
- Gene expression dysregulation, particularly involving microtubule and dynein-related genes, may compensate for CFAP161 loss, buffering the phenotype.
