Analysis of gene-trap mouse lines disrupting long intergenic non-coding RNA genes.

• Project/Area Number: 23310135
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Genome biology
• Research Institution: Kumamoto University
• Principal Investigator: ARAKI Kimi 熊本大学, 生命資源研究・支援センター, 教授 (90211705)
• Co-Investigator(Kenkyū-buntansha): ARAKI Masatake 熊本大学, 生命資源研究・支援センター (80271609)
• Project Period (FY): 2011-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥20,410,000 (Direct Cost: ¥15,700,000、Indirect Cost: ¥4,710,000); Fiscal Year 2014: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2013: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2012: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2011: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000)
• Keywords: 遺伝子トラップ / 非コードRNA / X-gal 染色 / 変異マウス / X-gal染色

Outline of Final Research Achievements

Many numbers of large intergenic non-coding RNAs (lincRNAs) has been reported, however, in vivo functions of lincRNAs have remained unclear. We have performed gene-trap insertional mutagenesis in mouse ES cells and isolated 1,067 trap lines. Among them, 33 clones have the trap vector integrated into lincRNA genes. We selected 13 clones and established the mouse lines. This study intended to analyze the function of lincRNAs in vivo using the gene-trap mice. We analyzed the expression patterns during development and adult mouse tissues by X-gal staining.Twelve trap lines have stained positively in various tissues,and many of lincRNAs were expressed in the brain. We produced homozygous mice through heterozygous intercrossing, and observed whether the homozygotes were born in the expected Mendelian ratio and whether they can survive until weaning. Although 12 trap lines have no apparent abnormalities, one line showed low birth rate of homozygotes suggesting abnormal development.

Research Products

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  • Peer Reviewed / Acknowledgement Compliant
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• [Journal Article] NRBF2 Regulates Autophagy and Prevents Liver Injury by Modulating Atg14L-Linked Class III Phosphatidylinositol-3 Kinase Activity. (2014)
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• [Journal Article] Disruption of Mbd5 in mice causes neuronal functional deficits and neurobehavioral abnormalities consistent with 2q23.1 microdeletion syndrome. (2014)
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• [Journal Article] Conformational change in transfer RNA is an early indicator of acute cellular damage (2014)
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• [Journal Article] Role of blood ribosomal protein S19 in coagulum resorption: a study using Gln137Glu-ribosomal protein S19 gene knock-in mouse. (2014)
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• [Journal Article] Disruption of actin-binding domain-containing Dystonin protein causes dystonia musculorum in mice. (2014)
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• [Journal Article] The kick-in system: a novel rapid knock-in strategy. (2014)
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• [Journal Article] Generation and analysis of serine protease inhibitor kazal type 3-cre driver mice. (2014)
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  • Journal Title: Exp. Anim. Volume: 63 Pages: 45-53
  • NAID: 130003391605
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• [Journal Article] Screening Methods to Identify TALEN-Mediated Knockout Mice. (2014)
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  • Journal Title: Exp. Anim. Volume: 63 Pages: 79-84
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• [Journal Article] Autophagy regulation in the pancreatic acinar cells is independent of epidermal growth factor receptor signaling. (2014)
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  • Journal Title: Biochem. Biophysic. Res. Comm. Volume: 446 Issue: 1 Pages: 224-230
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• [Journal Article] Loss of mTOR complex 1 induces developmental blockage in early T-lymphopoiesis and eradicates T-cell acute lymphoblastic leukemia cells. (2013)
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• [Journal Article] Perivascular adipose tissue-secreted angiopoietin-like protein2 (Angptl2) accelerates neointimal hyperplasia after endovascular injury. (2013)
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  • Journal Title: J Mol Cell Cardiol Volume: 57 Pages: 1-12
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  • Peer Reviewed
• [Journal Article] Gene-trap mutagenesis using Mol/MSM-1 embryonic stem cells from MSM/Ms mice (2013)
  • Author(s): Nakahara, M., Tateyama, H., Araki, M., Nakagata, N., Yamamura, K. and Araki, K.
  • Journal Title: Mammal Genome Volume: 24 Issue: 5-6 Pages: 228-239
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  • Peer Reviewed
• [Journal Article] Interaction of CD99 with its paralog CD99L2 positively regulates CD99L2 trafficking to cell surfaces. (2013)
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  • Journal Title: J. Immunol. Volume: 191 Issue: 11 Pages: 5730-5742
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• [Journal Article] HOT1 is a mammalian direct telomere repeat-binding protein contributing to telomerase recruitment. (2013)
  • Author(s): Kappei, D., Butter, F., Benda, C., Scheibe, M., Draskovic, I., Stevense, M., Novo, C.L., Basquin, C., Araki, M., Araki, K., Krastev, D.B., Kittler, R., Jessberger, R., Londono-Vallejo, J.A., Mann, M. and Buchholz, F.
  • Journal Title: EMBO J. Volume: 32 Issue: 12 Pages: 1681-1701
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• [Journal Article] TASK-2: a K2P K(+) channel with complex regulation and diverse physiological functions. (2013)
  • Author(s): Cid, L. P., Roa-Rojas, H. A., Niemeyer, M. I., Gonzalez, W., Araki, M., Araki, K. and Sepulveda, F. V.
  • Journal Title: Front Physiol. Volume: 4 Pages: 198-198
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• [Journal Article] Applications of cryopreserved unfertilized mouse oocytes for in vitro fertilization. (2013)
  • Author(s): Nakagata, N., Takeo, T., Fukumoto, K., Kondo, T., Haruguchi, Y., Takeshita, Y., Nakamuta, Y.,, Matsunaga, H., Tsuchiyama, S., Ishizuka, Y. and Araki, K.
  • Journal Title: Cryobiology. Volume: 67 Issue: 2 Pages: 188-192
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• [Journal Article] VMAT2 identified as a regulator of late-stage beta cell differentiation. (2013)
  • Author(s): Sakano, D., Shiraki, N., Kikawa, K., Yamazoe, T., Kataoka, M., Umeda, K., Araki, K., Mao, D., Matsumoto, S., Nakagata, N., Andersson, O., Stainier, D., Endo, F., Kume, K., Uesugi, M. and Kume, S.
  • Journal Title: Nat. Chem. Biol. Volume: 10 Issue: 2 Pages: 141-148
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• [Journal Article] Gene trap mutagenesis using Mol/MSM-1 ES cells established from MSM/Ms strain. (2013)
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  • Journal Title: Mamm. Genome. Volume: in press
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• [Journal Article] Ectopic expression of Ptf1a induces spinal defects, urogenital defects and anorectal malformations in Danforth's short tail mice. (2012)
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  • Journal Title: PLoS Genet. Volume: Vol.9 Issue: 2 Pages: 1-17
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• [Journal Article] mTORC1is essential for leukemia propagation but not stem cell self-renewal. (2012)
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  • Journal Title: J Clin Invest. Volume: 122 Issue: 6 Pages: 2114-2129
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• [Journal Article] A Cre Knock-in Mouse Line on the Sickle Tail Locus Induces Recombination in the Notochord and Intervertebral Discs. (2012)
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  • Journal Title: Genesis Volume: 50 Issue: 10 Pages: 758-765
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• [Journal Article] Macrophage-derived Angiopoietin- like Protein 2 Accelerates the Development of Abdominal Aortic Aneurysm. (2012)
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  • Journal Title: Arterioscler Thromb Vasc Biol Volume: 32(6) Issue: 6 Pages: 1400-1409
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• [Journal Article] CD99-clependent expansion of myeloid-derived suppressor cells and attenuation of graft-versus-host disease (2012)
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  • Journal Title: Mol.Cells Volume: 33 Issue: 3 Pages: 259-267
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• [Journal Article] mTORC1 is essential for leukemia-propagation but not stem cell self-renewal (2012)
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• [Journal Article] IGSF4 is a novel TCR zeta-chain-interacting protein that enhances TCR-mediated signaling (2011)
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• [Journal Article] Angiopoietin-like protein 2 is an important facilitator of inflammatory carcinogenesis and metastasis (2011)
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• [Journal Article] Nifedipine Increases Energy Expenditure by Increasing PGC-1α Expression in Skeletal Muscle (2011)
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  • Journal Title: Hypertension Research Volume: 34 Issue: 11 Pages: 1221-1227
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• [Journal Article] Essential roles of androgen signaling in wolffian duct stabilization and epididymal cell differentiation (2011)
  • Author(s): Murashima, A.
  • Journal Title: Endocrinology Volume: 152 Issue: 4 Pages: 1640-1651
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• [Journal Article] NSrp70 is a novel nuclear speckle-related protein that modulates alternative pre-mRNA splicing in vivo (2011)
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• [Journal Article] The cell cycle regulator Cdh1 controls the pool sizes of hematopoietic stem cells and mature lineage progenitors by protecting from genotoxic stress (2011)
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• [Presentation] CRISPR/Cas9による2本鎖切断・1本鎖切断を利用した場合のマウスES細胞における相同組換え効率の比較 (2014)
  • Author(s): 荒木喜美,牟田真由美,仙波 圭,竹田直樹,仁木大輔,松本 健,武田伊世,山村研一,大村谷昌樹,荒木正健
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 神奈川県（パシフィコ横浜）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] オートファジー不全と慢性膵炎発症メカニズムの解明 (2014)
  • Author(s): 大村谷昌樹,坂田和也,高城克暢,松元美由紀,荒木喜美,山村研一
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 神奈川県（パシフィコ横浜）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 椎間板変性モデルマウス(Danforth’s short tail)の発症原因の解明 (2014)
  • Author(s): 仙波 圭,荒木正健,池川志郎,飯田有俊,山村研一,荒木喜美
  • Organizer: 第29回日本整形外科学会基礎学術集会
  • Place of Presentation: 鹿児島県（城山観光ホテル）
  • Year and Date: 2014-10-09 – 2014-10-10
• [Presentation] 可変型遺伝子トラップマウスラインのパスウェイ及びジーンオントロジー解析 (2014)
  • Author(s): 荒木正健,中原 舞,中潟直己,山村研一,吉信公美子,荒木喜美
  • Organizer: 日本遺伝学会第86回大会
  • Place of Presentation: 滋賀県（長浜バイオ大学）
  • Year and Date: 2014-09-17 – 2014-09-19
• [Presentation] MSM/Ms由来ES細胞を用いたc-MafトランスジェニックMSM/Msマウスの作製と解析 (2014)
  • Author(s): 昇地高雅,舘山浩紀,高橋 智,山村研一,荒木喜美
  • Organizer: 日本遺伝学会第86回大会
  • Place of Presentation: 滋賀県（長浜バイオ大学）
  • Year and Date: 2014-09-17 – 2014-09-19
• [Presentation] 遺伝子トラップマウスを用いたlincRNAの生体内機能解析 (2014)
  • Author(s): 宮家幹子,森田彩香,柳井千佳,山添史雅,中原 舞,荒木美幸,岡本頼幸,伊東春香,大西雄一朗,國場訓山村研一,吉信公美子,荒木正健,荒木喜美
  • Organizer: 第28回モロシヌス研究会
  • Place of Presentation: 静岡県（国立遺伝学研究所）
  • Year and Date: 2014-06-27 – 2014-06-28
• [Presentation] Analysis of gene-trap mouse lines trapping long non-coding RNA genes.(Part1) (2013)
  • Author(s): 柳井千佳,山添史雅,中原 舞,宮家幹子,森田彩香,荒木美幸,岡本瀬幸,伊東春香,山村研一,吉信久美子,荒木正健,荒木喜美
  • Organizer: 第27回モロシヌス研究会
  • Place of Presentation: つくば市 理研BRC
• [Presentation] Analysis of gene-trap mouse lines trapping long non-coding RNA genes.(Part2) (2013)
  • Author(s): 山添史雅,柳井千佳,中原 舞,宮家幹子,森田彩香,荒木美幸,岡本瀬幸,伊東春香,山村研一,吉信久美子,荒木正健,荒木喜美
  • Organizer: 第27回モロシヌス研究会
  • Place of Presentation: つくば市 理研BRC
• [Presentation] 可変型遺伝子トラップ法を用いた骨代謝疾患に関与する新規遺伝子の探索 (2013)
  • Author(s): 関本朝久、黒木修司、舩元太郎、大田智美、中村志保子、濱田浩朗、山村研一、中原舞、荒木喜美、荒木正健、帖佐悦男
  • Organizer: 第86回日本整形外科学会学術総会
  • Place of Presentation: 広島 広島グリーンアリーナ
• [Presentation] 可変型遺伝子トラップ法を用いたLima1/EPLINの骨代謝における機能解析 (2013)
  • Author(s): 黒木修司、関本朝久、舩元太郎、大田智美、中村志保子、山村研一、中原舞、荒木喜美、荒木正健、帖佐悦男
  • Organizer: 第28回日本整形外科学会基礎学術集会
  • Place of Presentation: 千葉 幕張メッセ
• [Presentation] 遺伝子トラップマウスを用いたlincＲＮＡの機能解析 (2013)
  • Author(s): 中原 舞,山添史雅,柳井千佳,吉信公美子,山村研一,荒木正健,荒木喜美
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 兵庫 神戸ポートアイランド
• [Presentation] c-Ｍaf遺伝子導入ＭＳＭ/Ｍsマウスの作出と解析 (2013)
  • Author(s): 昇地高雅,荒木喜美,舘山浩紀,山村研一,高橋 智
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 兵庫 神戸ポートアイランド
• [Presentation] 可変型遺伝子トラップクローンの進展 (2013)
  • Author(s): 吉信公美子,來海葉子,慶田貴子,古閑成美,中原 舞,山村研一,荒木喜美,荒木正健
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 兵庫 神戸ポートアイランド
• [Presentation] ヘルパーＥＳ細胞を用いることによりキメラ作製困難なＥＳ細胞から生殖系列キメラを得る手法の開発 (2012)
  • Author(s): 荒木喜美
  • Organizer: 第５９回日本実験動物学会総会
  • Place of Presentation: 別府国際コンベンションセンター
• [Presentation] ＥＧＴＣマウスラインのGene Ontology解析 (2012)
  • Author(s): 荒木正健
  • Organizer: 第５９回日本実験動物学会総会
  • Place of Presentation: 別府国際コンベンションセンター
• [Presentation] 可変型遺伝子トラップ法を用いた骨代謝疾患に関与する新規遺伝子の探索 (2012)
  • Author(s): 黒木修司
  • Organizer: 第30回日本骨代謝学会学術集会
  • Place of Presentation: 東京 京王プラザホテル
• [Presentation] 可変型遺伝子トラップクローンデータベース（EGTC） (2012)
  • Author(s): 荒木正健
  • Organizer: 日本遺伝学会第84会大会
  • Place of Presentation: 九州大学医学部
• [Presentation] 変型遺伝子トラップ法を用いたLima1/EPLINの骨代謝における機能解析 (2012)
  • Author(s): 舩元太郎
  • Organizer: 第27回日本整形外科学会基礎学術集会
  • Place of Presentation: 名古屋国際会議場
• [Presentation] Danforth’s short tail (Sd) 変異の原因はPtf1aの異所性発現である (2012)
  • Author(s): 荒木喜美
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場
• [Presentation] 未知遺伝子をトラップした可変型遺伝子トラップマウスラインの解析 (2012)
  • Author(s): 吉信久美子
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場
• [Presentation] 可変型相同組換え法によるTtrおよびRbp遺伝子座のダブルヒト化マウスの作製 (2011)
  • Author(s): 李正花
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜)
  • Year and Date: 2011-12-13
• [Presentation] 新しい部位特異的組換えシステムDre/roxを用いたES細胞及びマウス個体での組換え効率の検討 (2011)
  • Author(s): 荒木喜美
  • Organizer: 第25回モロシヌス研究会
  • Place of Presentation: ベルナティオ(新潟)
  • Year and Date: 2011-07-08
• [Presentation] MSM/Msマウス由来ES細胞を用いた遺伝子トラップクローンの解析 (2011)
  • Author(s): 中原舞
  • Organizer: 第25回モロシヌス研究会
  • Place of Presentation: ベルナティオ(新潟)
  • Year and Date: 2011-07-08
• [Presentation] MSM由来ES細胞を用いた遺伝子操作マウスの作出と解析 (2011)
  • Author(s): 荒木喜美
  • Organizer: 第58回日本実験動物学会総会
  • Place of Presentation: タワーホール船橋(東京)(招待講演)
  • Year and Date: 2011-05-25
• [Presentation] EGTCマウスラインのGene Ontology解析 (2011)
  • Author(s): 荒木正健
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜)
• [Presentation] 可変型遺伝子トラップマウスにおけるプロモーター発現解析 (2011)
  • Author(s): 吉信公美子
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜)
• [Book] Genetic manipulations using Cre and mutant loxP sites. (ed. Alexei Morozov), in Controlled genetic manipulations. Neuromethods. Vol 65 (2012)
  • Author(s): Araki, K.
  • Publisher: Humana Press.
• [Remarks] 疾患モデル分野ホームページ
  • URL: http://irda-genetics.kuma-u.jp/index.html
• [Remarks] EGTC
  • URL: http://egtc.jp
• [Patent(Industrial Property Rights)] モデル動等動物の作出方法及びモデル動物 (2012)
  • Inventor(s): 大村谷 昌樹、荒木 喜美
  • Industrial Property Rights Holder: 国立大学法人熊本大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2012-06-09
• [Patent(Industrial Property Rights)] マウス系統を樹立する方法 (2011)
  • Inventor(s): 荒木正健、荒木喜美
  • Industrial Property Rights Holder: 国立大学法人熊本大学
  • Industrial Property Number: 2010-194910
  • Filing Date: 2011-08-30
  • Overseas
• [Patent(Industrial Property Rights)] マウス系統を樹立する方法 (2010)
  • Inventor(s): 荒木正健、荒木喜美
  • Industrial Property Rights Holder: 国立大学法人熊本大学
  • Industrial Property Number: 2010-194910
  • Filing Date: 2010-08-31