Identification and characterization of transcripts transmitted from sperm to oocyte

2010 Fiscal Year Final Research Report

• Project/Area Number: 20770145
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Molecular biology
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: KAWANO Mitsuoki The Institute of Physical and Chemical Research, LSA要素技術開発ユニット, 研究員 (00455338)
• Project Period (FY): 2008 – 2010
• Keywords: 精子RNA / 生殖細胞特異的小分子RNA / 世代伝達性RNA / 核局在モチーフ配列

Research Abstract

I report the discovery by deep sequencing technology of a suite of small noncoding RNAs in mature mouse sperm. Two of them were highly enriched in sperm and detectable exclusively in germ cells. Designated 'spR-12 and -13' for 'sperm RNAs'. After fertilization, they were maintained in low copy numbers in the egg and the dividing blastomeres. A strict nuclear localization was evidenced by microinjection of fluorescence-tagged RNAs into the male pronucleus of one-cell embryos. Stably maintained during the preimplantation period, they were equally distributed between blastomeres at cell division, remaining strictly localized in the nucleus until the blastocyst stage at which they were not detectable. A sequence motif UGGG(G)CGGG common to the two spRs was sufficient for nuclear addressing of unrelated small RNAs. A knock-down experiment performed by microinjection of the spR-12 and spR-13 complementary sequences linked to the nuclear addressing motif showed a deleterious effect suggestive of a functional role of the two RNAs in early embryogenesis.

Research Products

• [Journal Article] miRNA profiling of bilateral rat hippocampal CA3 by deep sequencing. (2011)
  • Author(s): Shinohara, Y., Yahagi, K., Kawano, M., Nishiyori, H., Kawazu, C., Suzuki, N., Manabe, R., Hirase, H.
  • Journal Title: Biochem. Biophys.Res.Commun. 409 Pages: 293-298
  • Peer Reviewed
• [Journal Article] Reduction of non-insert sequence reads by "dimer eliminator" LNA oligonucleotide for small RNA deep sequencing. (2010)
  • Author(s): Kawano, M., Kawazu, C., Lizio, M., Kawaji, H., Carninci, P., Suzuki, H., Hayashizaki, Y.
  • Journal Title: Biotechniques. 49(4) Pages: 751-755
  • Peer Reviewed
• [Journal Article] Efficient small RNA library preparation protocol. (2010)
  • Author(s): Kawano, M., Kawazu, C., Lizio, M., Kawaji, H., Carninci, P., Suzuki, H., Hayashizaki, Y.
  • Journal Title: Biotechniques Protocol Guide 2011 Pages: 4-6
• [Journal Article] Cross-mapping and the identification of editing sites in mature microRNAs in high-throughput sequencing libraries. (2010)
  • Author(s): de Hoon, M. J., Taft, R.J., Hashimoto, T., Kanamori-Katayama, M., Kawaji, H., Kawano, M., Kishima, M., Lassmann, T., Faulkner, J., Mattick, J. S., Daub, C. O., Carninci, P., Kawai, J., Suzuki, H., Hayashizaki, Y.
  • Journal Title: Genome Res. 20(2) Pages: 257-264
  • Peer Reviewed
• [Presentation] Reduction of non-insert sequence reads by LNA oligonucleotide for small RNA deep sequencing (2010)
  • Author(s): 川野光興、河津知佳、リッチオ・マリーナ、川路英哉、甲斐田薫、荒川貴博、カルニンチ・ピエロ、鈴木治和、林崎良英
  • Organizer: 第33回日本分子生物学会、第83回日本生化学会大会合同大会(BMB2010)
  • Place of Presentation: 神戸
  • Year and Date: 20101207-20101210
• [Presentation] 次世代小分子RNAネットワーク研究基盤の開発 (2009)
  • Author(s): 川野光興、河津知佳(発表者)、リッチオ・マリーナ、川路英哉、林崎良英
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20091209-20091212
• [Presentation] マウス初期胚の核に局在する精子特異的マイクロRNA様小分子RNA (2009)
  • Author(s): 川野光興、ラソールザデガン・ミノー、川路英哉、林崎良英
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20091209-20091212
• [Presentation] マウス初期胚の核に局在する精子特異的miRNA-like small RNA (2009)
  • Author(s): 川野光興、ラソールザデガン・ミノー、川路英哉、林崎良英
  • Organizer: 第11回日本RNA学会年会
  • Place of Presentation: 新潟
  • Year and Date: 20090727-20090729
• [Presentation] 世代を越えて機能するRNA の同定に向けて (2008)
  • Author(s): 川野光興、川路英哉、林崎良英
  • Organizer: 第31回日本分子生物学会・第81回日本生化学会大会合同大会(BMB2008)
  • Place of Presentation: 神戸
  • Year and Date: 20081209-20081212
• [Presentation] 精子中のRNA:世代を越えて機能するRNAの同定に向けて (2008)
  • Author(s): 川野光興、川路英哉、林崎良英
  • Organizer: 第10回日本RNA学会年会
  • Year and Date: 20080723-20080725
• [Presentation] RNA in sperm : Towards an identification of RNAs transmitting genetic information to zygote. (2008)
  • Author(s): Ma Kawano, M., Kawaji, H., Hasegawa, A., Severin, J., Sano, H., Hayashizaki, Y.
  • Organizer: Joint 7th Human Genome Organization (HUGO)-Pacific Meeting and the 8th Asia-Pacific Conference on Human Genetics, Symposium : Transmissible Nucleic Acids
  • Place of Presentation: Cebu, Philippines
  • Year and Date: 20080402-20080405
• [Presentation] RNA in sperm : Towards an identification of RNAs transmitted from sperm to zygote on fertilization. (2008)
  • Author(s): Kawano, M., Hasegawa, A., Sano, H., Kawaji, H., Severin, J., Lassmann, T., Hayashizaki, Y.
  • Organizer: Miami Winter Symposium Regulatory RNA in Biology and Human Health,
  • Place of Presentation: Miami, USA
  • Year and Date: 20080202-20080206
• [Remarks] ホームページ等
  • URL: http://genome.gsc.riken.jp/osc/members/Mitsuoki_Kawano.html