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Generation of Human Neurons by microRNA-Mediated Direct Conversion of Dermal Fibroblasts

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Nuclear Reprogramming

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2239))

Abstract

MicroRNAs (miRNAs), miR-9/9*, and miR-124 (miR-9/9*-124) display fate-reprogramming activities when ectopically expressed in human fibroblasts by erasing the fibroblast identity and evoking a pan-neuronal state. In contrast to induced pluripotent stem cell-derived neurons, miRNA-induced neurons (miNs) retain the biological age of the starting fibroblasts through direct fate conversion and thus provide a human neuron-based platform to study cellular properties inherent in aged neurons and model adult-onset neurodegenerative disorders using patient-derived cells. Furthermore, expression of neuronal subtype-specific transcription factors in conjunction with miR-9/9*-124 guides the miNs to distinct neuronal fates, a feature critical for modeling disorders that affect specific neuronal subtypes. Here, we describe the miR-9/9*-124-based neuronal reprogramming protocols for the generation of several disease-relevant neuronal subtypes: striatal medium spiny neurons, cortical neurons, and spinal cord motor neurons.

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Acknowledgments

The authors thank Shawei Chen for his inputs related to the protocol. V.A.C. is supported by a fellowship from the William N. and Bernice E. Bumpus Foundation. This work was supported by grants from NIH (RF1AG056296 and R01NS107488), CHDI foundation, Farrell Fund, and Collaborative Center for X-linked Dystonia Parkinsonism (CC-XDP) to A.S.Y.

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Author notes

  1. Victoria A. Church, Kitra Cates, Lucia Capano, Shivani Aryal and Woo Kyung Kim contributed equally to this work.

Authors and Affiliations

  1. Department of Developmental Biology, Center for Regenerative Medicine, Washington University School of Medicine, St. Louis, MO, USA

    Victoria A. Church, Kitra Cates, Lucia Capano, Shivani Aryal, Woo Kyung Kim & Andrew S. Yoo

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  1. Victoria A. Church
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  2. Kitra Cates
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  3. Lucia Capano
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  4. Shivani Aryal
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  5. Woo Kyung Kim
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  6. Andrew S. Yoo
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Corresponding author

Correspondence to Andrew S. Yoo .

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Editors and Affiliations

  1. Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL, USA

    Kejin Hu

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Church, V.A., Cates, K., Capano, L., Aryal, S., Kim, W.K., Yoo, A.S. (2021). Generation of Human Neurons by microRNA-Mediated Direct Conversion of Dermal Fibroblasts. In: Hu, K. (eds) Nuclear Reprogramming. Methods in Molecular Biology, vol 2239. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1084-8_6

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  • DOI: https://doi.org/10.1007/978-1-0716-1084-8_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1083-1

  • Online ISBN: 978-1-0716-1084-8

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