Identification of the miRNA targetome in hippocampal neurons using RIP-seq.

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Identification of the miRNA targetome in hippocampal neurons using RIP-seq. / Malmevik, Josephine; Petri, Rebecca; Klüssendorf, Thies; Knauff, Pina; Åkerblom, Malin; Johansson, Jenny G; Soneji, Shamit; Jakobsson, Johan.

In: Scientific Reports, Vol. 5, 12609, 2015.

Research output: Contribution to journalArticle

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TY - JOUR

T1 - Identification of the miRNA targetome in hippocampal neurons using RIP-seq.

AU - Malmevik, Josephine

AU - Petri, Rebecca

AU - Klüssendorf, Thies

AU - Knauff, Pina

AU - Åkerblom, Malin

AU - Johansson, Jenny G

AU - Soneji, Shamit

AU - Jakobsson, Johan

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

U2 - 10.1038/srep12609

DO - 10.1038/srep12609

M3 - Article

C2 - 26219083

VL - 5

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 12609

ER -