Development of technique for digital counting of DNA copy number at the single-cell level
Project/Area Number |
24360344
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Research Category |
Grant-in-Aid for Scientific Research (B)
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Allocation Type | Partial Multi-year Fund |
Section | 一般 |
Research Field |
Biofunction/Bioprocess
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Research Institution | Waseda University |
Principal Investigator |
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Co-Investigator(Kenkyū-buntansha) |
YOKOTA Yasuko 国立感染症研究所, 免疫部, 室長 (20182701)
HOSOKAWA Masahito 早稲田大学, ナノ・ライフ創新研究機構, 次席研究員(研究院助教) (60722981)
|
Project Period (FY) |
2012-04-01 – 2015-03-31
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Project Status |
Completed (Fiscal Year 2014)
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Budget Amount *help |
¥18,980,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥4,380,000)
Fiscal Year 2014: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2013: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2012: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
|
Keywords | DNA / HIV / デジタルカウント / 一分子PCR / 単一細胞 / デジタルカウンティング / 1分子PCR / マイクロ流体デバイス |
Outline of Final Research Achievements |
In this study, we aimed to develop a technique for digital counting of low copy viral DNA within infected cells. We developed a microfluidic device for generation of microdroplets for droplet-based digital PCR. In this method, the target genes were separately amplified within picoliter droplets and detected with fluorescent microscopy. The number of fluorescent droplets correlated with the copy numbers of target genes. In addition, we constructed lentivirus vector modeled after HIV. After optimization of the reaction conditions of droplet digital PCR, we demonstrated its highly sensitive detection of proviral DNA within a small number of infected cells during early infection process.
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Report
(4 results)
Research Products
(26 results)
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[Journal Article] Humanized mice dually challenged with R5 and X4 HIV-1 show preferential R5 viremia and restricted X4 infection of CCR5+CD4+ T cells.2015
Author(s)
Terahara, K., Ishige, M., Ikeno, S., Okada, S., Kobayashi-Ishihara, M., Ato, M., and Tsunetsugu-Yokota, Y
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Journal Title
Microbes and Infection
Volume: 10.1016/j.micinf.2015.02.002
Issue: 5
Pages: 378-386
DOI
Related Report
Peer Reviewed
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[Journal Article] Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR2013
Author(s)
Shota Ikeno, Moto-omi Suzuki, Mahmod Muhsen, Masayuki Ishige, Mie Kobayashi, Shinji Ohno, Makoto Takeda, Tetsuo Nakayama, Yuko Morikawa, Kazutaka Terahara, Seiji Okada, Haruko Takeyama, Yasuko Tsunetsugu-Yokota
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Journal Title
Frontiers in Microbiology
Volume: 4
Pages: 298-298
DOI
Related Report
Peer Reviewed
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[Journal Article] HIV-1 infection ex vivoaccelerates measles virus infection by upregulating signaling lymphocytic activation molecule (SLAM) in CD4+T cells2012
Author(s)
Mitsuki Y, Terahara K, Shibusawa K, Yamamoto T, Tsuchiya T, Ishige M, Kobayashi K, Morikawa Y, Nakayama T, Takeda M, Yanagi Y, Tsunetsugu-Yokota Y
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Journal Title
J Virol
Volume: 86
Issue: 13
Pages: 7227-7234
DOI
Related Report
Peer Reviewed
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