Abstract
The plasma membrane of living cells is an interface of material transfers and an antenna for outer signals. This chapter provides a guide on how to fabricate bioluminescent capsules for illuminating intracellular signaling and cargo protein delivery. The capsule consists of four components, which are, in consecutive order: a secretion peptide (SP), a host luciferase body (leader), a guest protein or peptide (cargo), and a membrane localization signal (MLS). Any guest protein, including a luciferase or a fluorescent protein, may be sandwiched between the host luciferase body and MLS and may be deliverable to the plasma membrane (PM), where the capsule waits for outer signals and to quickly release the embedded luciferase in response to a specific signal. The present strategy provides an efficient molecular vehicle for cargo proteins and imaging of intracellular molecular events in living cells without substrate-derived demerits of luciferases.
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Acknowledgments
This work was supported by a Grant-in-Aid for Young Scientists (B) from the Japan Society for the Promotion of Science (JSPS) [Grant number 23750096] and by a Grant-in-Aid for an A-Step program from the Japan Science and Technology Agency (JST) [Grant number AS232Z02075F].
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Kim, S.B. (2014). Fabrication of Bioluminescent Capsules and Live-Cell Imaging. In: Badr, C. (eds) Bioluminescent Imaging. Methods in Molecular Biology, vol 1098. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-718-1_10
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DOI: https://doi.org/10.1007/978-1-62703-718-1_10
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Publisher Name: Humana Press, Totowa, NJ
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