Function of promoter-associated non-coding RNA for the epigenome formation in the early mouse embryo

• Project/Area Number: 25660250
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Integrative animal science
• Research Institution: Kyushu University
• Principal Investigator: IMAMURA TAKUYA 九州大学, 医学(系)研究科(研究院), 助教 (90390682)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ノンコーディングRNA / DNAメチル化 / DNA脱メチル化 / 遺伝子活性化 / 人為操作胚 / 雌性発生胚 / 初期胚 / リプログラミング / ノンコーディング / RNA

Outline of Final Research Achievements

In mice, zygotic activation occurs for a wide variety of genes, mainly at the 2-cell stage. In this study, directional RNA-seq of MII oocytes and 2-cell embryos identified more than 1000 divergently transcribed long noncoding RNA (lncRNA)/mRNA gene pairs. Expression of these bidirectional promoter-associated noncoding RNAs (pancRNAs) was strongly associated with the upregulation of their cognate genes. Conversely, knockdown of three abundant pancRNAs led to reduced mRNA expression, accompanied by sustained DNA methylation even in the presence of enzymes responsible for DNA demethylation. In particular, microinjection of siRNA against the abundant pancRNA partner of interleukin 17d (Il17d) mRNA at the 1-cell stage caused embryonic lethality, which was rescued by supplying IL17D protein in vitro at the 4-cell stage. Thus, this novel class of lncRNAs can modulate the epigenetic machinery in cis to activate zygotic genes and is essential for preimplantation development.

Research Products

• [Journal Article] Gene activation-associated long noncoding RNAs function in mouse preimplantation development (2015)
  • Author(s): Hamazaki N, Uesaka M, Nakashima K, Agata K, Imamura T
  • Journal Title: Development Volume: 142 Pages: 910-920
  • DOI: 10.1242/dev.116996
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification of hypothalamic arcuate nucleus-specific enhancer region of kiss1 gene in mice. (2015)
  • Author(s): Goto, T., Tomikawa, J., Ikegami, K., Minabe, S., Abe, H., Fukanuma, T., Imamura, T., Takase, K., Sanbo, M., Tomita, K., Hirabayashi, M., Maeda, K., Tsukamura, H., Uenoyama, Y.
  • Journal Title: Mol Endocrinol Volume: 29 Issue: 1 Pages: 121-129
  • DOI: 10.1210/me.2014-1289
  • Peer Reviewed / Open Access
• [Journal Article] Epigenetic setting and reprogramming for neural cell fate determination and differentiation. (2014)
  • Author(s): Takuya Imamura, Masahiro Uesaka, Kinichi Nakashima
  • Journal Title: Philosophical transactions of the Royal Society of London. Series B Volume: 369 Issue: 1652 Pages: 1652-1652
  • DOI: 10.1098/rstb.2013.0511
  • Peer Reviewed
• [Journal Article] Bidirectional promoters are the major source of gene activation-associated non-coding RNAs in mammals. (2014)
  • Author(s): Uesaka M, Nishimura O, Go Y, Nakashima K, Agata K, Imamura T.
  • Journal Title: BMC Genomics Volume: 15 Issue: 1 Pages: 35-35
  • DOI: 10.1186/1471-2164-15-35
  • Peer Reviewed
• [Presentation] Bidirectional promoter-derived antisense noncoding RNA epigenetically regulates irreversible differentiation of PC12 cells (2015)
  • Author(s): Naoki Yamamoto, Kiyokazu Agata, Kinichi Nakashima, Takuya Imamura
  • Organizer: 第8回神経発生討論会
  • Place of Presentation: 九州大学病院キャンパス（福岡市）
  • Year and Date: 2015-03-19 – 2015-03-20
• [Presentation] Species-specific repertories of promoter-associated non-coding RNAs may contribute to the diversification of gene expression profile (2015)
  • Author(s): Masahiro Uesaka, Kiyokazu Agata, Kinichi Nakashima, Takuya Imamura
  • Organizer: 第8回神経発生討論会
  • Place of Presentation: 九州大学病院キャンパス（福岡市）
  • Year and Date: 2015-03-19 – 2015-03-20
• [Presentation] 長鎖ncRNAによるほ乳類エピゲノム制御 (2014)
  • Author(s): 今村拓也
  • Organizer: 第157回日本獣医学会学術集会
  • Place of Presentation: 北海道大学高等教育推進機構（札幌市）
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] マウス初期胚の全能性を支える遺伝子を活性化するノンコーディングRNAの同定 (2014)
  • Author(s): 濵崎伸彦, 上坂将弘, 中島欽一, 阿形清和, 今村拓也
  • Organizer: 第107回日本繁殖生物学会大会
  • Place of Presentation: 帯広畜産大学（帯広市）
  • Year and Date: 2014-08-21 – 2014-08-24
• [Presentation] Potential role for species-specific promoter-associated non-coding RNAs in the diversification of mammalian brain (2014)
  • Author(s): Masahiro Uesaka, Osamu Nishimura, Kinichi Nakashima, Kiyokazu Agata, Takuya Imamura
  • Organizer: 第47回日本発生生物学会年会
  • Place of Presentation: ウインクあいち（名古屋市）
  • Year and Date: 2014-05-27 – 2014-05-30
• [Presentation] Promoter-associated noncoding RNAs mediate gene-specific DNA demethylation for mouse preimplantation development (2014)
  • Author(s): Nobuhiko Hamazaki, Masahiro Uesaka, Kinichi Nakashima, Kiyokazu Agata, Takuya Imamura
  • Organizer: 第8回日本エピジェネティクス研究会年会
  • Place of Presentation: 伊藤国際学術研究センター（東京都文京区）
  • Year and Date: 2014-05-25 – 2014-05-27
• [Presentation] PC12細胞分化過程におけるエピジェネティック制御に関わる promoter-associated noncoding RNAの機能 (2014)
  • Author(s): 山本直樹、上坂将弘、中島欽一、阿形清和、今村拓也
  • Organizer: 第8回日本エピジェネティクス研究会年会
  • Place of Presentation: 伊藤国際学術研究センター（東京都文京区）
  • Year and Date: 2014-05-25
• [Presentation] Promoter-associated noncoding RNAs mediate gene-specific DNA demethylation for mouse preimplantation development (2013)
  • Author(s): Nobuhiko Hamazaki, Masahiro Uesaka, Kinichi Nakashima, Kiyokazu Agata, Takuya Imamura
  • Organizer: 第七回エピジェネティクス研究会
  • Place of Presentation: 奈良県新公会堂（奈良市）
• [Presentation] Mammalian epigenome formation mediated by promoter-associated noncoding RNA (2013)
  • Author(s): Masahiro Uesaka, Osamu Nishimura, Kinichi Nakashima, Kiyokazu Agata, Takuya Imamura
  • Organizer: 第七回エピジェネティクス研究会
  • Place of Presentation: 奈良県新公会堂（奈良市）
• [Presentation] マウス受精胚におけるnoncoding RNAを介した遺伝子プロモーターのエピジェネティック変換 (2013)
  • Author(s): 浜崎伸彦, 上坂将弘, 阿形清和, 今村拓也
  • Organizer: 第106回日本繁殖生物学会
  • Place of Presentation: 東京農工大学（府中市）
• [Book] Roles of Epigenetics in the Neural Stem Cell and Neuron: In: J. Peedicayil, D. Grayson, D. Avramopoulos (eds), ISBN 9780124171145 Epigenetics in Psychiatry (2014)
  • Author(s): Naoki Yamamoto, Masahiro Uesaka, Takuya Imamura, Kinichi Nakashima
  • Total Pages: 664
  • Publisher: Elsevier
• [Remarks] 九大プレスリリース
  • URL: http://www.kyushu-u.ac.jp/pressrelease/2015/2015_02_05.pdf
• [Remarks] 九大医学研究院HP
  • URL: http://www.med.kyushu-u.ac.jp/app/modules/information/detail.php?i=753&c=10
• [Remarks] 九大医学系学府HP
  • URL: http://www.grad.med.kyushu-u.ac.jp/researchinfo/detail.php?i=753
• [Remarks] 九州大学医学研究院基盤幹細胞学分野
  • URL: http://www.scb.med.kyushu-u.ac.jp/imamura/