Generation of mice models of AML/MDS with large genomic deletion using CRISPR/Cas9 genome editing

• Project/Area Number: 26860735
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Hematology
• Research Institution: Hiroshima University
• Principal Investigator: Nagamachi Akiko 広島大学, 原爆放射線医科学研究所, 助教 (20585153)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ゲノム編集 / 骨髄異型性症候群 / MDS / 骨髄異形成症候群 / -7/7q-

Outline of Final Research Achievements

We use the CRISPR/Cas9 system of genome editing to generate model mice of myeloid malignancy which is deleted the candidate 7q- responsible genes, Miki, CG-NAP, and Samd9L. By injection of circular plasmid expressing sgRNA and hCas9 into mouse zygotes, we generated mice carried the mutations at the target locus with high efficiency. However, it was difficult to generate the multiple genes (Miki/CG-NAP/Samd9L) deficient mice by co-injection of the plasmids targeting a single allele. Thus we are trying to generate multiple gene deficient mice use a different technique to complement the current knockout project.

Research Products

• [Journal Article] Involvement of resistin-like molecule β in the development of methionine-choline deficient diet-induced non-alcoholic steatohepatitis in mice. (2016)
  • Author(s): Okubo H, Kushiyama A, Sakoda H, Nakatsu Y, Iizuka M, Taki N, Fujishiro M, Fukushima T, Kamata H, Nagamachi A, Inaba T, Nishimura F, Katagiri H, Asahara T, Yoshida Y, Chonan O, Encinas J, Asano T.
  • Journal Title: Sci Rep. Volume: 6 Issue: 1 Pages: 20157-20157
  • DOI: 10.1038/srep20157
  • NAID: 120006796541
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] -7/7q- syndrome in myeloid-lineage hematopoietic malignancies: attempts to understand this complex disease entity. (2015)
  • Author(s): Honda H, Nagamachi A, Inaba T.
  • Journal Title: Oncogene Volume: 34 Issue: 19 Pages: 2413-2425
  • DOI: 10.1038/onc.2014.196
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] SETBP1 mutations drive leukemic transformation in ASXL1-mutated MDS. (2015)
  • Author(s): Inoue D, Kitaura J, Matsui H, Hou HA, Chou WC, Nagamachi A, Kawabata KC, Togami K, Nagase R, Horikawa S, Saika M, Micol JB, Hayashi Y, Harada Y, Harada H, Inaba T, Tien HF, Abdel-Wahab O, Kitamura T.
  • Journal Title: Leukemia Volume: 29(4) Issue: 4 Pages: 847-57
  • DOI: 10.1038/leu.2014.301
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Fbxl10 overexpression in murine hematopoietic stemcells induces leukemia involving metabolic activation and upregulation of Nsg2 (2015)
  • Author(s): Ueda T, Nagamachi A, Takubo K, Yamasaki N, Matsui H, Kanai A, Nakata Y, Ikeda K Konuma T, Oda H, Wolff L, Honda Z, Wu X, Helin K, Iwama A, Suda T, Inaba T, Honda H.
  • Journal Title: Blood Volume: 125 Issue: 22 Pages: 3437-3446
  • DOI: 10.1182/blood-2014-03-562694
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] Low Miki expression in myelodysplastic syndromes results in polynuclear cell formation through decondensation of chromosomes in prolonged prometaphase. (2015)
  • Author(s): Nagamachi A., Ozaki Y., Matsui H., Kanai A., Inaba T.
  • Organizer: The 57th American Society of Hematology Annual Meeting
  • Place of Presentation: Orlando, USA
  • Year and Date: 2015-12-06
  • Int'l Joint Research
• [Presentation] eEF1A2 is a target gene of DNA demethylating agents for improving anemia of MDS (2015)
  • Author(s): 長町安希子、松井啓隆、金井昭教、稲葉俊哉
  • Organizer: 第77回日本血液学会学術総会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-17
• [Presentation] Exploring haplo-insufficieny of multiple anti-oncogenes that facilitate malignant transformation (2015)
  • Author(s): 長町安希子、稲葉俊哉
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2015-10-09
• [Presentation] 低線量率放射線照射による染色体異常出現頻度の解析 (2015)
  • Author(s): 長町安希子, 藤岡来実, 稲葉俊哉, 神谷研二.
  • Organizer: 第56回原子爆弾後障害研究会
  • Place of Presentation: 広島
  • Year and Date: 2015-06-07
• [Presentation] Roles of translational machinery in epigenetic therapy in MDS (2014)
  • Author(s): 長町安希子
  • Organizer: 第76回日本血液学会学術総会
  • Place of Presentation: 大阪国際会議場
  • Year and Date: 2014-10-13 – 2014-10-15
• [Presentation] Roles of translational machinery in epigenetic therapy in MDS (2014)
  • Author(s): 長町安希子
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] 7q欠失責任遺伝子候補Samd9Lマウスモデルの作製と機能解析 (2014)
  • Author(s): 長町安希子
  • Organizer: 第10回麒麟塾
  • Place of Presentation: コクヨホール
  • Year and Date: 2014-07-12
• [Presentation] ユビキチン修飾酵素A20が造血機構に与える影響 (2014)
  • Author(s): 長町安希子
  • Organizer: 第55回原子爆弾後障害研究会
  • Place of Presentation: 長崎原爆資料館
  • Year and Date: 2014-06-01
• [Presentation] モノソミー7を伴うAML/MDSのモデルマウスの作製 (2014)
  • Author(s): 長町安希子
  • Organizer: 「細胞運命制御」領域 若手研究者の会
  • Place of Presentation: 浜名湖グランドホテル
  • Year and Date: 2014-04-18 – 2014-04-19