 Hi, I am Sudeep Theroy and I am an associate professor at the Institute of Molecular and Cell Biology in Singapore. My lab focuses on the differentiation and function of cilia. Cilia-tiny hair-like organelles present on the surface of most eukaryotic cells. Cilia can be motile such as those found in our respiratory tracts that beat continuously to clear mucus or imotile and dedicated to transmit sensory signals such as those found in our eyes and nose for a sense of sight and smell. A few years ago, through genetic studies in the zebrafish embryo, we discovered that the transcription factor FoxG1 is the master regulator of motile and cilia differentiation. More recently, we performed a large-scale functional genomic screen for the targets of FoxG1 and discovered the CCDC-11 gene, which encodes a coil-coil-containing protein as one of the targets of FoxG1. Hi, I am Vijay Narsimhar and I am a post-doctor fellow with Sudeep Theroy. Together with my colleagues Shuba and Sudeep, we studied the function of CCDC-11 in cilia using genetic analysis in zebrafish embryos. We first determined the expression of CCDC-11 using in situ aberration technique and found that the gene was expressed in tissues that differentiate motile cilia. CCDC-11 was expressed in the kidney tube use, throat plate or the spinal canal and the KV or kufa vesicles. KV is analogous to the ventral load of the mammars and beating cilia in these structures drives a signaling cascade that determines the laterality of the visceral organs. We used micro-injection technique to introduce morpholinos into zebrafish embryos that enables the function of CCDC-11 gene in zebrafish. We found that when CCDC-11 activity was compromised in zebrafish embryos, various phenotypes that are characteristics of defects in the motile cilia were observed, namely, curl-body axes, otter defects, and prominent left flight asymmetry defects were observed in the embryos. We contacted our collaborator, Prof. Ahmed Omran, to investigate the role of CCDC-11 as a potential candidate gene for primary seniority dyskinesia. Hi, I'm Ahmed. I'm actually the head of a research lab that is very much interested in primary cilia dyskinesia and I'm a pediatrician caring for patients with this kind of rare lung disease. Hi, I'm Ray Mehtaed and I'm a PhD student in Prof. Omran's lab. Because we were informed by Sudip Duroy that there is a candidate gene, CCDC-11, that is likely to be a good gene responsible for primary cilia dyskinesia, we investigated our total genome scans of several consanguinous families and identified in this family on chromosome 18 an area of homozygosity by descent. And RIM went ahead and tried to test whether this gene might be related to the disease. Okay, indeed, we have found in one of these patients a number of P1069 a homozygous nonsense mutation changing a cytosentrify mean resulting in a clodostope and predicting an early termination of the protein. The 12-year-old male actually suffered from mild respiratory symptoms like chronic rhinitis and some cough, and he also displayed cytosin versus so you see that the heart is actually on the right and the stomach is actually also on the right. So we have a complete mirror of image. In order to confirm the loss of function mutation we have identified, we have studied the localization of the protein CCDC-11, performing immunofluorescence analysis and using antibodies targeting the CCDC-11 protein. And we have found in control cells that the CCDC-11 has an axonimol localization. It is localized to hold the whole axonimol length. And in the respiratory epithelial cells of the patient, we have found that no CCDC-11 was detectable in the sedia. And this confirms the loss of function mutation we have identified. In order to discern the defect code by CCDC-11 deficiency, we analyzed cross-sections by transmission electron microscopy of CCDC-11 deficiency in the sedia and we have found no observable ultra-structural defect. And consistently, when performing immunofluorescence analysis, studying the localization of outer dine-in-arm proteins, inner dine-in-arm proteins or nexine dine-in regulatory complexes proteins, we have found that all of these proteins localize normally in CCDC-11 mutant sedia. Immunofluorescence analysis shows that zebrafish CCDC-11 protein localizes to the axonimol and the base of the kidney sedia, which is very similar to the localization of CCDC-11 protein in human respiratory sedia. However, the protein localized only to the base of the KV sedia and also to the base of the spinal cord sedia. To evaluate the effect of CCDC-11 noctore on the motility of sedia and zebrafish, we performed live imaging of wild-type and CCDC-11 morphine-injected embryos. We found that the motility of the sedia in the kidney of CCDC-11 morphines were only marginally impaired when compared to the wild-type sedia. However, we found out that the rotational motion of sedia in the KV and the flow plate were strongly impaired. We performed transmission electron microscopy to determine the effect of loss of CCDC-11 on the ultrastructure of KV sedia. On the ultrastructure level, there appeared to be loss of outer dining arm from the CCDC-11 deficient KV sedia. Our findings demonstrate that CCDC-11 has a conserved essential role in sedia of the vertebrate left-right organiser. To the best of our knowledge, this is the first sediary component which has a differential localization and function in different kinds of motile sedia. Thank you and happy reading.