The DOI matches its Crossref record; 31 of the 31 references it cites were found in a registry or an index.
Consensus guided mutagenesis of Renilla luciferase yields enhanced stability and light output
Open accessAbstract
Luciferases, which have seen expansive employment as reporter genes in biological research, could also be used in applications where the protein itself is conjugated to ligands to create probes that are appropriate for use in small animal imaging. As the bioluminescence activity of commonly used luciferases is too labile in serum to permit this application, specific mutations of Renilla luciferase, selected using a consensus sequence driven strategy, were screened for their ability to confer stability of activity in serum as well as their light output. Using this information, a total of eight favorable mutations were combined to generate a mutant Renilla luciferase (RLuc8) that, compared with the parental enzyme, is 200-fold more resistant to inactivation in murine serum and exhibits a 4-fold improvement in light output. Results of the mutational analysis were also used to generate a double mutant optimized for use as a reporter gene. The double mutant had half the resistance to inactivation in serum of the native enzyme while yielding a 5-fold improvement in light output. These variants of Renilla luciferase, which exhibit significantly improved properties compared with the native enzyme, will allow enhanced sensitivity in existing luciferase-based assays as well as enable the development of novel probes labeled with the luciferase protein.
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Couturier, C., & Deprez, B. (2012) · Frontiers in Endocrinology · doi:10.3389/fendo.2012.00100 · Cited by 52
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Bungart, B. L., Dong, L., Sobek, D., Sun, G. Y., Yao, G., & Lee, J. C.-M. (2014) · Nanomedicine: Nanotechnology, Biology and Medicine · doi:10.1016/j.nano.2013.10.007 · Cited by 27
Granberg, K. L., Sakamaki, S., Larsson, N., Bergström, F., Fuchigami, R., Niwa, Y., Ryberg, E., Backmark, A., Kato, H., Miyazaki, S., et al. (2024) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.3c02184 · Cited by 26
Beyond luciferase-luciferin system: modification, improved imaging and biomedical application
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Metadata
- Authors
- Loening, A. M.
- Venue
- Protein Engineering Design and Selection
- Publisher
- Oxford University Press (OUP)
- Published
- 2006-07-20
- Volume
- 19
- Issue
- 9
- Pages
- 391-400
- ISSN
- 0269-2139 (ISSN-L)
- 1741-0126
- 1741-0134
- OpenAlex
- W2098956821
- Cited by
- 417
- References
- 31
- FWCI
- 7.12
- Type
- journal article
- Language
- en
- Open access
- bronze
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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