2011 Fiscal Year Final Research Report
In vivo Regulation of the Cell Fates Utilizing a Novel Inducible System of Protein Expression
| Project/Area Number |
21591231
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hematology
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| Research Institution | The University of Tokyo |
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Principal Investigator |
OTSU Makoto 東京大学, 医科学研究所, 特任准教授 (30361330)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAUCHI Yasuo 北里大学, 医学部, 講師 (60286359)
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| Project Period (FY) |
2009 – 2011
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| Keywords | 造血幹細胞 / レトロウイルスベクター / レンチウイルスベクター / 発現調節 / 細胞老化 |
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| Research Abstract |
As planned, I constructed the inducible retroviral system that could allow rapid and dose-related regulation of expression of the target protein. It, however, turned out that the degradation-domain(DD)/Shield-1 system did have an intrinsic defect in itself, which was compatible with significant expression leak. I thus decided to change gears to the combined use of DD/Shield-1 and tetracycline-inducible system, the latter of which was originally developed by Dr. Bujard. I utilized so-called "All-in-One LV", the lentiviral system that was generated by my colleague to allow containing all the required elements for the system to work within a single cassette. Among the combination tested, I finally figured out that the system comprised of Tet-ON and DD/Shield-1 was the best performer so that it allowed the lowest leak and prompt expression upon dual stimulation. Although the overall progress remain behind the schedule due to the change of plans, I believe that the system is now ready to be tested in vivo, once the inducible target gene is introduced within this newly developed dual-regulation lentiviral vector.
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[Journal Article] Frequent pathway mutations of splicing machinery in myelodysplasia2011
Author(s)
Yoshida K, Sanada M, Shiraishi Y, Nowak D, Nagata Y, Yamamoto R, Sato Y, Sato-Otsubo A, Kon A, Nagasaki M, Chalkidis G, Suzuki Y, Shiosaka M, Kawahata R, Yamaguchi T, Otsu M, Obara N, Sakata-Yanagimoto M, Ishiyama K, Mori H, Nolte F, Hofmann WK, Miyawaki S, Sugano S, Haferlach C, Koeffler HP, Shih LY, Haferlach T, Chiba S, Nakauchi H, Miyano S, Ogawa S
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Journal Title
Nature
Volume: 478
Pages: 64-69
Peer Reviewed
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[Journal Article] Heterozygous ITGA2B R995W mutation inducing constitutive activation of the alphaIIbbeta3 receptor affects proplatelet formation and causes congenital macrothrombocytopenia2011
Author(s)
Kunishima S, Kashiwagi H, Otsu M, Takayama N, Eto K, Onodera M, Miyajima Y, Takamatsu Y, Suzumiya J, Matsubara K, Tomiyama Y, Saito H
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Journal Title
Blood
Volume: 117
Pages: 5479-5484
Peer Reviewed
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[Journal Article] Transient activation of c-MYC expression is critical for efficient platelet generation from human induced pluripotent stem cells2010
Author(s)
Takayama N, Nishimura S, Nakamura S, Shimizu T, Ohnishi R, Endo H, Yamaguchi T, Otsu M, Nishimura K, Nakanishi M, Sawaguchi A, Nagai R, Takahashi K, Yamanaka S, Nakauchi H, Eto K
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Journal Title
J Exp Med
Volume: 207
Pages: 2817-30
Peer Reviewed
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[Journal Article] Reduction of N-glycolylneuraminic acid in human induced pluripotent stem cells generated or cultured under feeder-and serum-free defined conditions2010
Author(s)
Hayashi Y, Chan T, Warashina M, Fukuda M, Ariizumi T, Okabayashi K, Takayama N, Otsu M, Eto K, Furue MK, Michiue T, Ohnuma K, Nakauchi H, Asashima M
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Journal Title
PLoS One
Volume: 5
Pages: e14099
Peer Reviewed
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[Journal Article] Gain-of-function of mutated C-CBL tumour suppressor in myeloid neoplasms2009
Author(s)
Sanada M, Suzuki T, Shih LY, Otsu M, Kato M, Yamazaki S, Tamura A, Honda H, Sakata-Yanagimoto M, Kumano K, Oda H, Yamagata T, Takita J, Gotoh N, Nakazaki K, Kawamata N, Onodera M, Nobuyoshi M, Hayashi Y, Harada H, Kurokawa M, Chiba S, Mori H, Ozawa K, Omine M, Hirai H, Nakauchi H, Koeffler HP, Ogawa S
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Journal Title
Nature
Volume: 460
Pages: 904-8
Peer Reviewed
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