Functional Analysis of MicroRNAs in Neurogenesis During Mouse Cortical Development
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The advantage of using in utero electroporation is that it can study the gene function during neurodevelopment in vivo. Using functional analysis of a microRNA (miRNA) gene as an example, this protocol describes a set of techniques that are crucial for the success of neurogenesis studies, including mice time mating, plasmid preparation, utero electroporation following miRNA injection into mice embryonic brain ventricle, labeling of proliferating cells with EDU (ethynyldeoxyuridine), cryosectioning, immunofluorescence staining, and confocal microscopic analysis. This chapter also provides detailed technical tips regarding experimental planning, mouse surgery, multi-embryo injection with different plasmids, electroporation, and maintenance of pregnant mother with post-electroporated embryo.
Key wordsIn utero electroporation MicroRNA Cerebral cortex Immunohistochemistry Neural proliferation Neurogenesis Cortical development
We thank Dr. Paul Kim and Ms. Neo Yan Zhuang for their technical support. This research was supported by the Singapore Ministry of Health’s National Medical Research Council grant to W. Z and Translational Clinical Research grant to L.Z.
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