RNAi-mediated knockdown of mouse melanocortin-4 receptor in vitro and in vivo, using an siRNA expression construct based on the mir-187 precursor
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- Kato Minoru
- Research Unit/Neuroscience, Mitsubishi Tanabe Pharma Corporation, 1000 Kamoshida, Aoba-ku, Yokohama, Kanagawa 227-0033, Japan
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- Huang Yi-Ying
- Laboratory of Chronogenomics, Mitsubishi Kagaku Institute of Life Science, 11 Minamiooya, Machida, Tokyo 194-8511, Japan
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- Matsuo Mina
- Experimental Animal Laboratory, Mitsubishi Kagaku Institute of Life Science, 11 Minamiooya, Machida, Tokyo 194-8511, Japan Present address: Division of Animal Experimental Laboratory, Life Science Research Center, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan
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- Takashina Yoko
- Pharmacology Laboratory, Mitsubishi Tanabe Pharma Corporation, 1000 Kamoshida, Aoba-ku, Yokohama, Kanagawa 227-0033, Japan
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- Sasaki Kazuyo
- Pharmacology Laboratory, Mitsubishi Tanabe Pharma Corporation, 1000 Kamoshida, Aoba-ku, Yokohama, Kanagawa 227-0033, Japan
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- Horai Yasushi
- Safety Research Laboratory, Mitsubishi Tanabe Pharma Corporation, 1000 Kamoshida, Aoba-ku, Yokohama, Kanagawa 227-0033, Japan
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- Juni Aya
- Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
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- Kamijo Shin-Ichi
- Experimental Animal Laboratory, Mitsubishi Kagaku Institute of Life Science, 11 Minamiooya, Machida, Tokyo 194-8511, Japan Present address: Division of Planning of Laboratory Animal Facility, Lifewill Corporation, 2740-12 Shimo-oyamada, Machida, Tokyo 194-0202, Japan
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- Saigo Kaoru
- Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
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- Ui-Tei Kumiko
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
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- Tei Hajime
- Laboratory of Chronogenomics, Mitsubishi Kagaku Institute of Life Science, 11 Minamiooya, Machida, Tokyo 194-8511, Japan Present address: Graduate School of Natural Science and Technology, Kanazawa University, Kakuma, Kanazawa, Ishikawa 920-1192, Japan
書誌事項
- タイトル別名
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- RNAi-mediated knockdown of mouse melanocortin-4 receptor <i>in vitro</i> and <i>in vivo</i>, using an siRNA expression construct based on the mir-187 precursor
この論文をさがす
説明
<p>RNA interference (RNAi) is a powerful tool for the study of gene function in mammalian systems, including transgenic mice. Here, we report a gene knockdown system based on the human mir-187 precursor. We introduced small interfering RNA (siRNA) sequences against the mouse melanocortin-4 receptor (mMc4r) to alter the targeting of miR-187. The siRNA-expressing cassette was placed under the control of the cytomegalovirus (CMV) early enhancer/chicken β-actin promoter. In vitro, the construct efficiently knocked down the gene expression of a co-transfected mMc4r-expression vector in cultured mammalian cells. Using this construct, we generated a transgenic mouse line which exhibited partial but significant knockdown of mMc4r mRNA in various brain regions. Northern blot analysis detected transgenic expression of mMc4r siRNA in these regions. Furthermore, the transgenic mice fed a normal diet ate 9% more and were 30% heavier than wild-type sibs. They also developed hyperinsulinemia and fatty liver as do mMc4r knockout mice. We determined that this siRNA expression construct based on mir-187 is a practical and useful tool for gene functional studies in vitro as well as in vivo.</p>
収録刊行物
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- Experimental Animals
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Experimental Animals 66 (1), 41-50, 2017
公益社団法人 日本実験動物学会
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キーワード
詳細情報 詳細情報について
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- CRID
- 1390001205044762624
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- NII論文ID
- 130005298582
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- NII書誌ID
- AA11032321
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- ISSN
- 18817122
- 00075124
- 13411357
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- NDL書誌ID
- 027861987
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- PubMed
- 27725374
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
- KAKEN
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- 抄録ライセンスフラグ
- 使用不可