Optogenetic Control of Subcellular Protein Location and Signaling in Vertebrate Embryos.
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Buckley, C. (2019). Optogenetic Control of Subcellular Protein Location and Signaling in Vertebrate Embryos.. [Book chapter]. https://doi.org/10.1007/978-1-4939-9009-2_10
This chapter describes the use of optogenetic heterodimerization in single cells within whole-vertebrate embryos. This method allows the use of light to reversibly bind together an "anchor" protein and a "bait" protein. Proteins can therefore be directed to specific subcellular compartments, altering biological processes such as cell polarity and signaling. I detail methods for achieving transient expression of fusion proteins encoding the phytochrome heterodimerization system in early zebrafish embryos (Buckley et al., Dev Cell 36(1):117-126, 2016) and describe the imaging parameters used to achieve subcellular light patterning.
Animals, Vertebrates, Zebrafish, RNA, Messenger, Microinjections, Signal Transduction, Gene Expression, Protein Transport, Embryonic Development, Transgenes, Plasmids, Optogenetics
Wellcome Trust (208758/Z/17/Z)
External DOI: https://doi.org/10.1007/978-1-4939-9009-2_10
This record's DOI: https://doi.org/10.17863/CAM.75309