Manipulation technology development of chicken pluripotent stem cells for constructing transgenic bioreactors

• Project/Area Number: 26289316
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Kyushu University
• Principal Investigator: Kamihira Masamichi 九州大学, 工学(系)研究科(研究院), 教授 (40202022)
• Co-Investigator(Renkei-kenkyūsha): KAWABE Yoshinori 九州大学, 工学研究院, 助教 (30448401)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,120,000 (Direct Cost: ¥12,400,000、Indirect Cost: ¥3,720,000); Fiscal Year 2016: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2014: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
• Keywords: バイオテクノロジー / 生物・生体工学 / 生体バイオリアクター / ニワトリ / 多能性幹細胞 / 応用動物

Outline of Final Research Achievements

This study aimed to develop a technology to generate transgenic chickens derived from pluripotent stem cells as a new production platform for biopharmaceuticals. Total RNAs extracted from chicken blastodermal cells, containing pluripotent stem cells, were applied for the massive sequencing technology to analyze expression profiles of reprogramming factor genes. Using reprogramming factor genes identified by the transcriptome analysis, chicken induced pluripotent stem cells were generated. Furthermore, a genome editing technique, CRISPR/Cas9 system was applied for targeting a sex determination gene, DMRT1. A targeting vector designed efficiently altered the target region in DMRT1 gene of chicken cells.

Research Products

• [Presentation] 遺伝子導入ニワトリ卵を用いた経口免疫ワクチンによるアレルギー治療法の開発 (2016)
  • Author(s): 大坪 嵩征, 小畑 玲奈, 河邉 佳典, 井藤 彰, 上平 正道
  • Organizer: 第53回化学関連支部合同九州大会
  • Place of Presentation: 北九州国際会議場
• [Presentation] 遺伝子導入ニワトリによるアレルギー治療のためのTGF-beta1の生産 (2015)
  • Author(s): 小畑 玲奈, 河邉 佳典, 井藤 彰, 上平 正道
  • Organizer: 第67回日本生物工学会大会
  • Place of Presentation: 城山観光ホテル（鹿児島）
  • Year and Date: 2015-10-28
• [Presentation] ニワトリ胚盤葉細胞の網羅的発現遺伝子解析による多能性幹細胞誘導因子の探索 (2015)
  • Author(s): 椎葉 温, 河邉 佳典, 福丸 詩帆, 山田 拓矢, 井藤 彰, 上平 正道
  • Organizer: 化学工学会第４７回秋季大会
  • Place of Presentation: 北海道大学札幌キャンパス
  • Year and Date: 2015-09-09
• [Presentation] ニワトリ胚盤葉細胞の網羅的遺伝子発現解析 (2015)
  • Author(s): 椎葉 温, 河邉 佳典, 福丸 詩帆, 山田 拓矢, 井藤 彰, 上平 正道
  • Organizer: 第52回化学関連支部合同九州大会
  • Place of Presentation: 北九州国際会議場
  • Year and Date: 2015-06-27
• [Presentation] Biopharmaceutical protein production using transgenic chickens (2015)
  • Author(s): Masamichi Kamihira
  • Organizer: 5th Annual International Conference on Advances in Biotechnology (BIOTECH2015)
  • Place of Presentation: Indian Institute of Technology, Kanpur, India
  • Year and Date: 2015-03-13
  • Invited
• [Presentation] Transcriptome analysis of chicken blastodermal cells (2014)
  • Author(s): Yoshinori Kawabe, Shiho Fukumaru, Nodoka Shiiba, Takuya Yamada, Akira Ito, Masamichi Kamihira
  • Organizer: The 27th Annual and International Meeting of Japanese Association for Animal Cell Technology (JAACT2014)
  • Place of Presentation: Kitakyushu International Conference Center (Kitakyushu)
  • Year and Date: 2014-11-14
• [Presentation] ニワトリ多能性幹細胞樹立のための胚盤葉細胞の網羅的遺伝子発現解析 (2014)
  • Author(s): 山田 拓矢, 福丸 詩帆, 椎葉 温, 河邉 佳典, 井藤 彰, 上平 正道
  • Organizer: 化学工学会第４６回秋季大会
  • Place of Presentation: 九州大学伊都キャンパス（福岡）
  • Year and Date: 2014-09-17
• [Remarks] 上平研究室ホームページ
  • URL: http://www.chem-eng.kyushu-u.ac.jp/lab3/
• [Remarks] 研究者情報
  • URL: http://hyoka.ofc.kyushu-u.ac.jp/search/details/K002690/index.html
• [Remarks] 研究室ホームページ
  • URL: http://www.chem-eng.kyushu-u.ac.jp/lab3/