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In utero gene transfer system for embryos before neural tube closure reveals a role for Hmga2 in the onset of neurogenesis

Naohiro Kuwayama, Yusuke Kishi, Yurie Maeda, Yurie Nishiumi, Yutaka Suzuki, Haruhiko Koseki, Yusuke Hirabayashi, Yukiko Gotoh
doi: https://doi.org/10.1101/2020.05.14.086330
Naohiro Kuwayama
1Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033,Japan
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Yusuke Kishi
1Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033,Japan
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  • For correspondence: ykisi@mol.f.u-tokyo.ac.jp
Yurie Maeda
1Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033,Japan
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Yurie Nishiumi
1Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033,Japan
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Yutaka Suzuki
2Department of Computational Biology, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo 277-8561, Japan
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Haruhiko Koseki
3Laboratory for Developmental Genetics, RIKEN Center for Integrative Medical Sciences (RIKEN-IMS), Tsurumi-ku, Yokohama 230-0045, Japan
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Yusuke Hirabayashi
4Graduate School of Engineering, The University of Tokyo, Tokyo 113-0033, Japan
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Yukiko Gotoh
1Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033,Japan
5International Research Center for Neurointelligence (WPI-IRCN), The University of Tokyo, Tokyo 113-0033, Japan
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Abstract

During the early stage of mammalian neural development, neuroepithelial cells (NECs) proliferate and increase their pool size before switching their fate to become neurogenic radial glial progenitors (RGPs). The timing of this expansion-to-neurogenic phase transition is strictly regulated so as to determine the proper number of progenitors and differentiated cell types that constitute the brain. The molecular mechanism underlying this switch has remained poorly understood, however, in part because of the difficulty associated with manipulation of gene expression in NECs before their transition to RGPs, which occurs before neural tube closure. We have now developed a simple and efficient method to manipulate gene expression in mouse neocortical NECs in the expansion phase by in utero injection of viral vectors at embryonic day 7.0 to 8.0. With the use of this method, we found that knockdown of the chromatin-associated protein Hmga2 in NECs inhibited the onset of the neurogenic phase in the neocortex. This effect of Hmga2 knockdown was accompanied by inhibition of the expression of a set of genes targeted by Polycomb group proteins, which repress neurogenic genes in cortical progenitors. Our study thus establishes a novel method for manipulation of gene expression in the early stage of mouse brain development as well as uncovers a key molecular player in the generation of neurogenic progenitors in the developing mouse neocortex.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted May 15, 2020.
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In utero gene transfer system for embryos before neural tube closure reveals a role for Hmga2 in the onset of neurogenesis
Naohiro Kuwayama, Yusuke Kishi, Yurie Maeda, Yurie Nishiumi, Yutaka Suzuki, Haruhiko Koseki, Yusuke Hirabayashi, Yukiko Gotoh
bioRxiv 2020.05.14.086330; doi: https://doi.org/10.1101/2020.05.14.086330
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In utero gene transfer system for embryos before neural tube closure reveals a role for Hmga2 in the onset of neurogenesis
Naohiro Kuwayama, Yusuke Kishi, Yurie Maeda, Yurie Nishiumi, Yutaka Suzuki, Haruhiko Koseki, Yusuke Hirabayashi, Yukiko Gotoh
bioRxiv 2020.05.14.086330; doi: https://doi.org/10.1101/2020.05.14.086330

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