title: Identification of the miRNA targetome in hippocampal neurons using RIP-seq.
abstract: MicroRNAs (miRNAs) are key players in the regulation of neuronal processes by targeting a large network of target messenger RNAs (mRNAs). However, the identity and function of mRNAs targeted by miRNAs in specific cells of the brain are largely unknown. Here, we established an adeno-associated viral vector (AAV)-based neuron-specific Argonaute2:GFP-RNA immunoprecipitation followed by high-throughput sequencing to analyse the regulatory role of miRNAs in mouse hippocampal neurons. Using this approach, we identified more than two thousand miRNA targets in hippocampal neurons, regulating essential neuronal features such as cell signalling, transcription and axon guidance. Furthermore, we found that stable inhibition of the highly expressed miR-124 and miR-125 in hippocampal neurons led to significant but distinct changes in the AGO2 binding of target mRNAs, resulting in subsequent upregulation of numerous miRNA target genes. These findings greatly enhance our understanding of the miRNA targetome in hippocampal neurons.
authors.affiliation: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, SE-221 84 Lund, Sweden.
authors.forename: Josephine
authors.initials: J
authors.lastname: Malmevik
authors.affiliation: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, SE-221 84 Lund, Sweden.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Petri
authors.affiliation: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, SE-221 84 Lund, Sweden.
authors.forename: Thies
authors.initials: T
authors.lastname: Klussendorf
authors.affiliation: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, SE-221 84 Lund, Sweden.
authors.forename: Pina
authors.initials: P
authors.lastname: Knauff
authors.affiliation: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, SE-221 84 Lund, Sweden.
authors.forename: Malin
authors.initials: M
authors.lastname: Åkerblom
authors.affiliation: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, SE-221 84 Lund, Sweden.
authors.forename: Jenny
authors.initials: J
authors.lastname: Johansson
authors.affiliation: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, SE-221 84 Lund, Sweden.
authors.forename: Shamit
authors.initials: S
authors.lastname: Soneji
authors.affiliation: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, SE-221 84 Lund, Sweden.
authors.forename: Johan
authors.initials: J
authors.lastname: Jakobsson
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000378:metabolism; D001369:Axons / Q000378:metabolism; D000229:Dependovirus / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics; D006624:Hippocampus / Q000378:metabolism*; D047468:Immunoprecipitation / Q000379:methods; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D009474:Neurons / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics
chemical_list: C526735:Ago2 protein, mouse; D060565:Argonaute Proteins; D035683:MicroRNAs; D012333:RNA, Messenger