Protocol for High-Content Screening for the Impact of Overexpressed MicroRNAs on Primary Motor Neurons
In this chapter we provide a protocol for the design and usage of automated, high-content microscopy screening that enables the investigation of microRNA (miRNA) impact on primary motor neuron. High-content screening (HCS) platforms facilitate superior precision in research and are scalable to study in parallel multiple genetic, molecular, or cellular conditions. miRNAs are critical for neuronal function and for brain integrity and are considered attractive candidate targets for therapy in many neuropathologies. Therefore, HCS platforms provide a novel paradigm for exploring the impact of miRNA expression, applicable for functional pathways discovery in an academic setting, or towards development of therapeutics in the pharma industry.
Keywords:High-content screening Primary motor neuron Automated microscopy Morphometry MicroRNA miRNAs
We thank Noga Kozer, Alexander Plotnikov, Haim M. Barr, Shir Katz, Hila Weiss, and Michal Eisenberg (Weizmann Institute of Science) for collaborations on developing the protocols. Raquel Fine and Cherill Banks for editing. This work was supported by grants from the ALS-Therapy Alliance, Israel Science Foundation, Legacy-heritage Fund, Bruno and Ilse Frick Foundation for Research on ALS, Yeda CEO fund, Minna-James-Heineman Stiftung through Minerva, ERC consolidator program, Nella and Leon Benoziyo Center for Neurological Diseases, Y. Leon Benoziyo Institute for Molecular Medicine, the Weizmann Institute of Science and A. Alfred Taubman via IsrALS, Kekst Family Institute for Medical Genetics, David and Fela Shapell Family Center for Genetic Disorders Research, Crown Human Genome Center, Yeda Sela Center, Helen and Martin Kimmel Institute for Stem Cell Research, the Maurice and Vivienne Wohl Biology Endowment, the Nathan, Shirley, Philip and Charlene Vener New Scientist Fund, Julius and Ray Charlestein Foundation, Celia Benattar Memorial Fund for Juvenile Diabetes, the Wolfson Family Charitable Trust, and Adelis Foundation. TY was supported by postdoctoral fellowship from Teva Pharmaceutical Industries Ltd as part of the Israeli National Network of Excellence in Neuroscience (NNE) and by the postdoctoral fellowship from the Sara Lee Schupf foundation. E.H. is Head of Nella and Leon Benoziyo Center for Neurological Diseases and the lab is further supported by Dr. Sydney Brenner and Friends.