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A Dual Promoter System to Monitor IFN-γ Signaling <i>in vivo</i> at Single-cell Resolution
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- Tanaka Taisei
- Laboratory of Bioimaging and Cell Signaling, Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University
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- Konishi Yoshinobu
- Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School
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- Ichise Hiroshi
- Lymphocyte Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health
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- Tsukiji Shinya
- Department of Nanopharmaceutical Sciences, Nagoya Institute of Technology
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- Matsuda Michiyuki
- Laboratory of Bioimaging and Cell Signaling, Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University Institute for Integrated Cell-Material Sciences, Kyoto University
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- Terai Kenta
- Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University
Bibliographic Information
- Other Title
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- A Dual Promoter System to Monitor IFN-γ Signaling in vivo at Single-cell Resolution
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Description
<p>IFN-γ secreted from immune cells exerts pleiotropic effects on tumor cells, including induction of immune checkpoint and antigen presentation, growth inhibition, and apoptosis induction. We combined a dual promoter system with an IFN-γ signaling responsive promoter to generate a reporter named the interferon sensing probe (ISP), which quantitates the response to IFN-γ by means of fluorescence and bioluminescence. The integration site effect of the transgene is compensated for by the PGK promoter-driven expression of a fluorescent protein. Among five potential IFN-γ-responsive elements, we found that the interferon γ-activated sequence (GAS) exhibited the best performance. When ISP-GAS was introduced into four cell lines and subjected to IFN-γ stimulation, dose-dependency was observed with an EC50 ranging from 0.2 to 0.9 ng/mL, indicating that ISP-GAS can be generally used as a sensitive biosensor of IFN-γ response. In a syngeneic transplantation model, the ISP-GAS-expressing cancer cells exhibited bioluminescence and fluorescence signals in an IFN-γ receptor-dependent manner. Thus, ISP-GAS could be used to quantitatively monitor the IFN-γ response both in vitro and in vivo.</p><p>Key words: in vivo imaging, tumor microenvironment, interferon-gamma, dual promoter system</p>
Journal
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- Cell Structure and Function
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Cell Structure and Function 46 (2), 103-111, 2021
Japan Society for Cell Biology
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Details 詳細情報について
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- CRID
- 1390290493077151744
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- NII Article ID
- 130008131966
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- ISSN
- 13473700
- 03867196
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- HANDLE
- 2433/268705
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- NDL BIB ID
- 032032645
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- PubMed
- 34744115
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- Text Lang
- en
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- Article Type
- journal article
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- Data Source
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- JaLC
- IRDB
- NDL Search
- Crossref
- PubMed
- CiNii Articles
- KAKEN
- OpenAIRE
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- Abstract License Flag
- Disallowed