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Quantitation of Transcription and Clonal Selection of Single Living Cells with β-Lactamase as Reporter
Abstract
Gene expression was visualized in single living mammalian cells with β-lactamase as a reporter that hydrolyzes a substrate loaded intracellularly as a membrane-permeant ester. Each enzyme molecule changed the fluorescence of many substrate molecules from green to blue by disrupting resonance energy transfer. This wavelength shift was detectable by eye or color film in individual cells containing less than 100 β-lactamase molecules. The robust change in emission ratio reveals quantitative heterogeneity in real-time gene expression, enables clonal selection by flow cytometry, and forms a basis for high-throughput screening of pharmaceutical candidate drugs in living mammalian cells.
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Cited by
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Liu, S.-Y., Aliyari, R., Chikere, K., Li, G., Marsden, M. D., Smith, J. K., Pernet, O., Guo, H., Nusbaum, R., Zack, J. A., et al. (2013) · Immunity · doi:10.1016/j.immuni.2012.11.005 · Cited by 678
HDACs, histone deacetylation and gene transcription: from molecular biology to cancer therapeutics
Gallinari, P., Marco, S. D., Jones, P., Pallaoro, M., & Steinkühler, C. (2007) · Cell Research · doi:10.1038/sj.cr.7310149 · Cited by 678
A sensitive and specific enzyme-based assay detecting HIV-1 virion fusion in primary T lymphocytes
Cavrois, M., de Noronha, C., & Greene, W. C. (2002) · Nature Biotechnology · doi:10.1038/nbt745 · Cited by 475
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Metadata
- Authors
- Zlokarnik, G., Negulescu, P. A., Knapp, T. E., Mere, L., Burres, N., Feng, L., Whitney, M., Roemer, K., & Tsien, R. Y.
- Venue
- Science
- Published
- 1998-01-02
- Volume
- 279
- Issue
- 5347
- Pages
- 84-88
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W2139855058
- Cited by
- 690
- References
- 0
- FWCI
- 30.64
- Type
- journal article
- Language
- en
- Open access
- closed
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- Zlokarnik, G., Negulescu, P. A., Knapp, T. E., Mere, L., Burres, N., Feng, L., Whitney, M., Roemer, K., & Tsien, R. Y. (1998). Quantitation of Transcription and Clonal Selection of Single Living Cells with β-Lactamase as Reporter. Science. https://doi.org/10.1126/science.279.5347.84
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
