A multiple gene expression switch using a multi-integrase system on a mammalian artificial chromosome vector.
| Project/Area Number |
25540134
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Life / Health / Medical informatics
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| Research Institution | Tottori University |
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Principal Investigator |
Ohbayashi Tetsuya 鳥取大学, 生命機能研究支援センター, 准教授 (80348804)
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| Co-Investigator(Renkei-kenkyūsha) |
Nakamura Kazuomi 鳥取大学, 生命機能研究支援センター, プロジェクト研究員 (90598137)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | 人工染色体ベクター / インテグレース / レポーターシステム / ゲノム編集 / ゲノム改変 / ゲノムプログラム / 遺伝子発現 |
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| Outline of Final Research Achievements |
Site-specific recombinases (SSR), such as Cre or Flp recombination target cassettes, have been successfully excised or inverted by a single SSR to regulate transgene expression. This study investigated the potential for expanding the multiple regulation of transgenes using three different integrase systems. We designed three excision cassettes that expressed luciferase, where the luciferase expression could be exchanged to a fluorescent protein by site-specific recombination. Each cassette that was regulated independently by a different integrase was connected in tandem and inserted into a mouse artificial chromosome (MAC) vector in CHO cells. By transient expression of integrase, the targeted luciferase activity was lost and fluorescence was activated. These results suggest that the combined use of these integrase systems in a defined locus on a MAC vector permits the multiple regulation of transgene expression and might contribute to genomic or cell engineering.
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Report
(4 results)
Research Products
(16 results)
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[Journal Article] Use of a Human Artificial Chromosome for Delivering Trophic Factors in a Rodent Model of Amyotrophic Lateral Sclerosis.2015
Author(s)
Watanabe Y, Kazuki Y, Kazuki K, Ebiki M, Nakanishi M, Nakamura K, Yoshida Yamakawa M, Hosokawa H, Ohbayashi T, Oshimura M, Nakashima K.
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Journal Title
Mol Ther Nucleic Acids.
Volume: 4
Pages: e253-e253
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Phenotype specific analyses reveal distinct regulatory mechanism for chronically activated p53.2015
Author(s)
Kirschner K, Samarajiwa SA, Cairns JM, Menon S, Pérez-Mancera PA, Tomimatsu K, Bermejo-Rodriguez C, Ito Y, Chandra T, Narita M, Lyons SK, Lynch AG, Kimura H, Ohbayashi T, Tavaré S, Narita M.
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Journal Title
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Exploring the origin and limitations of kidney regeneration.2015
Author(s)
Endo T, Nakamura J, Sato Y, Asada M, Yamada R, Takase M, Takaori K, Oguchi A, Iguchi T, Higashi AY, Ohbayashi T, Nakamura T, Muso E, Kimura T, Yanagita M.
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Journal Title
J Pathol.
Volume: 236(2)
Issue: 2
Pages: 251-63
DOI
Related Report
Peer Reviewed
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