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CSHL CELL & DEVELOPMENTAL BIOLOGY OF XENOPUS COURSE (2025-2029) - ABSTRACT IN VIVO ANIMAL MODELS ARE AN IMPORTANT TOOL FOR THE UNDERSTANDING OF HUMAN DEVELOPMENT AND DISEASE. STUDIES USING THE FROG XENOPUS HAVE MADE SIGNIFICANT CONTRIBUTIONS TO OUR UNDERSTANDING OF FUNDAMENTAL PROCESSES SUCH AS CELL CYCLE REGULATION, TRANSCRIPTION, TRANSLATION AND MANY OTHER TOPICS. XENOPUS IS REMARKABLE FOR STUDYING DEVELOPMENT AND DISEASE, INCLUDING BIRTH DEFECTS, CANCER, AND STEM CELL BIOLOGY. BECAUSE XENOPUS ARE EASY TO RAISE, PRODUCING MANY THOUSANDS OF EGGS PER DAY, THESE FROGS HAVE EMERGED AS A PREMIERE MODEL FOR UNDERSTANDING OF HUMAN BIOLOGY FROM THE FUNDAMENTAL BUILDING BLOCKS TO THE WHOLE ORGANISM. THE COLD SPRING HARBOR LABORATORY COURSE ON CELL & DEVELOPMENTAL BIOLOGY OF XENOPUS: GENE DISCOVERY & DISEASE PROVIDES EXTENSIVE LABORATORY EXPOSURE TO THE BIOLOGY AND MANIPULATION OF EMBRYOS FROM THE FROGS XENOPUS LAEVIS AND XENOPUS TROPICALIS. THE COURSE PREPARES STUDENTS FOR INDEPENDENT RESEARCH IN THE CELL, GENETIC, AND MOLECULAR ANALYSIS OF VERTEBRATE DEVELOPMENT. IT INTRODUCES THE UNIQUE ATTRIBUTES OF XENOPUS WHICH ALLOW THE FUNCTIONAL ANALYSIS OF DEVELOPMENTALLY IMPORTANT GENES, WHILST AT THE SAME TIME INTRODUCING THE MAJOR TECHNOLOGIES OF MODERN MOLECULAR AND CELLULAR DEVELOPMENTAL BIOLOGY, IN A SYSTEM THAT ALLOWS DIRECT MANIPULATION OF EMBRYOS AND VISUALIZATION OF DEVELOPMENTAL PROCESSES. AT THE END OF THE COURSE, STUDENTS ARE READY TO EMBARK ON CUTTING EDGE RESEARCH USING THIS POWERFUL MODEL OF HUMAN DEVELOPMENT. THE XENOPUS COURSE IS DESIGNED TO TRAIN STUDENTS HOW TO TAKE ADVANTAGE OF THESE UNIQUE QUALITIES. IT IS OFTEN DIFFICULT FOR INVESTIGATORS TO INITIATE STUDIES OF XENOPUS DEVELOPMENT ON THEIR OWN BECAUSE THE TECHNIQUES FOR RAISING EMBRYOS OR MANIPULATING THEM ARE UNFAMILIAR OR BECAUSE THEIR BACKGROUND KNOWLEDGE OF DEVELOPMENTAL BIOLOGY IS INADEQUATE. THIS COURSE IS DESIGNED TO PROVIDE STUDENTS WITH A CONCENTRATED AND REPEATED EXPOSURE TO THE MOST IMPORTANT RESEARCH TECHNIQUES, AND THE RESEARCHERS DOING THEM, SO THAT THEY INITIATE THEIR OWN INDEPENDENT RESEARCH PROJECTS AFTER RETURNING TO THEIR HOME INSTITUTIONS. THE COURSE PROVIDES AN INTENSIVE EXPOSURE, OVER A TWO-WEEK PERIOD, TO THE BIOLOGY AND MANIPULATION OF XENOPUS EMBRYOS. THE COURSE ALSO PROVIDES A RESOURCE NETWORK THAT THE STUDENTS CAN EASILY ACCESS AFTER THEY HAVE RETURNED TO THEIR HOME LABORATORIES. THROUGH THESE ROUTES, THE COURSE TRAINS INVESTIGATORS AT THE ADVANCED GRADUATE, POSTDOCTORAL AND FACULTY LEVELS TO USE XENOPUS IN THEIR RESEARCH DURING THE COURSE, THE STUDENTS ANALYZE ANY PHENOTYPES GENERATED FROM CRISPR/CAS9 BASED GENE DEPLETION WHILE LEARNING THE DIVERSE ARRAY OF TECHNIQUES AVAILABLE IN XENOPUS. IN PREVIOUS COURSES, WE HAVE GUIDED STUDENTS IN THE ABLATION OF A WIDE VARIETY OF GENES AND HELPED THEM DESIGN SUITABLE ASSAYS FOR THEIR BIOLOGICAL INTERESTS. MOST RECENTLY, STUDENTS HAVE TARGETED AUTISM GENES, THYROID GENES AND IMMUNE MODULATORS, SEVERAL OF WHICH HAVE ALREADY LED TO PUBLICATIONS. APPROACHES COVERED WILL INCLUDE MICROINJECTION AND MOLECULAR MANIPULATIONS SUCH AS CRISPR/CAS9 KNOCKOUTS, ANTISENSE MORPHOLINO-BASED DEPLETIONS, TRANSGENICS, AND MRNA OVEREXPRESSION. IN ADDITION, STUDENTS CAN COMBINE THESE TECHNIQUES WITH EXPLANT AND TRANSPLANT METHODS TO SIMPLIFY OR TEST TISSUE LEVEL INTERACTIONS. ADDITIONAL METHODS INCLUDE MRNA IN SITU HYBRIDIZATION, HYBRIDIZATION CHAIN REACTION (HCR), AND PROTEIN IMMUNOHISTOCHEMISTRY AS WELL AS BASIC BIOINFORMATIC TECHNIQUES FOR GENE COMPARISON AND FUNCTIONAL ANALYSIS. GENOMIC AND SYSTEMS BIOLOGY APPROACHES WILL BE INTRODUCED, INCLUDING SINGLE-CELL TECHNOLOGY. BIOCHEMICAL APPROACHES SUCH AS PROTEOMICS AND MASS SPECTROMETRY AND BIOMECHANICAL CONCEPTS WILL ALSO BE DISCUSSED. FINALLY, TO VISUALIZE SUBCELLULAR AND INTERCELLULAR ACTIVITIES, WE WILL INTRODUCE A VARIETY OF SAMPLE PREPARATION AND IMAGING METHODS INCLUDING TIME-LAPSE, FLUORESCENT IMAGING, OPTICAL COHERENCE TOMOGRAPHY AND CONFOCAL MICROSCOPY. IT IS ANTICIPATED THAT THE BASIC ORGANIZATION OF THE COURSE AND SOME