University of Crete 4 articles published in JoVE Genetics Using the GAL4-UAS System for Functional Genetics in Anopheles gambiae Beth Crawford Poulton1, Fraser Colman1, Amalia Anthousi1,2,3, Linda Grigoraki1, Adriana Adolfi1, Amy Lynd1, Gareth John Lycett1 1Department of Vector Biology, Liverpool School of Tropical Medicine, 2IMBB FORTH, 3Department of Biology, University of Crete The bipartite GAL4-UAS system is a versatile tool for modification of gene expression in a controlled spatiotemporal manner which permits functional genetic analysis in Anopheles gambiae. The procedures described for using this system are a semi-standardized cloning strategy, sexing and screening of pupae for fluorescent protein markers and embryo fixation. Biology Assessment of de novo Protein Synthesis Rates in Caenorhabditis elegans Margarita Elena Papandreou*1,2, Konstantinos Palikaras*1,2, Nektarios Tavernarakis1,2 1Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, Greece, 2Department of Basic Sciences, Faculty of Medicine, University of Crete, Heraklion, 70013, Crete, Greece Here, we introduce and describe a nonradioactive and noninvasive method to assess de novo protein synthesis in vivo, utilizing the nematode Caenorhabditis elegans and fluorescence recovery after photobleaching (FRAP). This method can be combined with genetic and/or pharmacological screens to identify novel modulators of protein synthesis. Biology Modeling Age-Associated Neurodegenerative Diseases in Caenorhabditis elegans Konstantinos Palikaras1,2, Nektarios Tavernarakis1,2 1Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, Greece, 2Department of Basic Sciences, Faculty of Medicine, University of Crete, Heraklion, 70013, Crete, Greece Here, we introduce and describe widely accessible methodologies utilizing some versatile nematode models, including hyperactivated ion channel-induced necrosis and protein aggregate-induced neurotoxicity, to monitor and dissect the cellular and molecular underpinnings of age-associated neurodegenerative diseases. Medicine In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice Evandro F. Fang1,6, Konstantinos Palikaras2, Nuo Sun3, Elayne M. Fivenson1, Ryan D. Spangler4, Jesse S. Kerr1, Stephanie A. Cordonnier1, Yujun Hou1, Eszter Dombi5, Henok Kassahun6, Nektarios Tavernarakis2,7, Joanna Poulton5, Hilde Nilsen6, Vilhelm A. Bohr1,8 1Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, 2Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology - Hellas, 3Center for Molecular Medicine, National Heart Lung and Blood Institute, National Institutes of Health, 4Laboratory of Neurosciences, National Institute on Aging, National Institutes of Health, 5Nuffield Department of Obstetrics and Gynaecology, University of Oxford, 6Department of Clinical Molecular Biology, University of Oslo and Akershus University Hospital, 7Department of Basic Sciences, Faculty of Medicine, University of Crete, 8Danish Center for Healthy Aging, University of Copenhagen Mitophagy, the process of clearing damaged mitochondria, is necessary for mitochondrial homeostasis and health maintenance. This article presents some of the latest mitophagy detection methods in human cells, Caenorhabditis elegans, and mice.
